Phytochemicals In Cervical Cancer Therapy: A Review of Recent Advances in Drug Design and Development

Abstract

Cervical cancer is a main cause of cancer-related death among women worldwide. Long-term infections with high-risk strains of the human papillomavirus (HPV), such as HPV-16 and HPV-18, are a major cause of cervical cancer, it is also major causes of cancer-related mortality among women globally. Side effects and resistance indicate the need for more effective and specific therapy, even with the availability of HPV vaccines and chemoradiotherapy. Alongside advancements in silico drug discovery techniques, this research investigates the therapeutic properties of phytoconstituents derived from medicinal plants. The article explains the molecular foundation of HPV carcinogenesis, including how the oncoproteins E6 and E7 in tumour suppressors such as p53 and Rb. The research on natural compounds present in several plants that are thought to have anti-cervical cancer properties. This activity is authenticated using molecular docking, gene expression profiling and studies that involve cell lines. Major phytochemicals like lycopodine, andrographolide, berberine, and rotundifuran can trigger apoptosis, stop proliferation, and affect crucial carcinogenic pathways. These results highlight the importance of natural substances as additional or alternative therapeutic agents and advocate for ongoing research on these compounds via bioinformatics and experimental verification.

Keywords

Introduction

Figure 1. Age-specific Incidence of cervical cancer worldwide in terms of the age

Structure And Function Of HPV

Human papillomavirus (HPV) infection is a major cause for cervical cancer (cc). Prophylactic vaccines were developed as a result of this  to prevent HPV infection and its assays . E1, E2, E4, E5, E6 are the six early proteins to which HPV codes. E7 proteins do reactivation of cell replication mechanism through the p^RB protein.⁶ Among risk strains of HPV, HPV 16 and 18 are closely associated with cervical cancer as well as a subdivision of cancer of the vulva, vagina.⁷ Early proteins of HPV 16, E6, and E7 play a crucial role in tumour formation.²  E6  oncoprotein has become centre of research activities across the world for the development of drug against the HPV positive cancer. The low risk of cervical cancer is presented by HPV 11, which usually causes benign hyperproliferative lesions as genital warts and oral papillomatosis ^1. Intervention of the tumour suppressant protein P53 and PRB together with viral oncoproteins such as E6 and E7 are responsible for cervical cancer. ^1&22 Studies show that E6 will bind to multiple ranges of cellular proteins playing functional role in cell cycle control and their regulations . Viral capsid of HPV is composed of 360 molecules of protein L1, which is the major capsid protein. Minor capsid L2 protein embedded inside the L1 proteins. The L1 and L2 are late genes that encode the structural proteins involved in the capsid formation. ⁹                                                 

Figure 2.   HPV Proteins and their function

L2 is the minor capsid protein that forms the wrapping of the HPV genome collectively with the L1 protein. Transportation of genomic DNA into the nucleus is primarily done by L2 proteins. E5 oncoproteins degranulate multiple cellular mediators such as EGF growth factors and their receptors. E5 interrupts the regulation of some epidermal growth factor receptors (EGFR) and signalling of other molecules which are involved in survival, growth, differentiation of the cell.  cervical cancer.¹ In India, 15 years and above, women are at the highest risk of developing cervical cancer. Human papillomavirus (HPV) is a DNA virus belonging to the family Papillomaviridae . The five genera of the 39 genera in the papillomaviridae family that are represented by HPVs are alpha, beta, gamma, mu, and nu. The alpha-papillomavirus are the one that causes genital warts.¹⁰

Figure 3. "Progression of HPV Infection to Cervical Cancer: Risk Factors and Precancerous Lesions"

It contains no envelope and the size ranges from 50 to 55 nm, consisting of a double-stranded DNA genome comprising 7200 to 8000 base pairs.⁷ It is estimated that there are 300 apparent genotypes of HPV, of which only 200 varieties have the capability of causing the disease in humans.⁴  WHO recommended a global eradication commencement of four cases per 100000 women years and the execution of a triple intervention strategy, comprising of vaccinating at least 90% of girls hostile to HPV by the age of 15 years.^3&7 Accessibility of highly effective natural therapeutics can be opted as alternative means to treat HPV and related diseases. Numerous plant-based compounds have been discovered as a prospective source for the development of cytotoxic agents for treatment and prevention of cervical cancer over the last few years.⁸ Medicinal plants such as “Allium sativum, Artemisia annua, Piper nigrum, Oxalis corniculate, Podophyllum hexandrum, Rheum emodi, Moringa oleifera’’ are some of Himalayan Herbs which are traditionally used as potential Inhibitory source of HPV in cervical cancer treatment ⁸. Naturally derived potential compounds from the plant source are the safest and effective therapeutics when compared with those of synthetically derived compounds. sourcing of the natural compounds from the botanical origin and their purification processes an expensive and time-consuming approach .¹¹ Drug discovery process as a prominent advantage of screening of potential bioactive compounds from the other molecules and interacting with the target molecular docking techniques as a major lead. Fresh milestones in computer-aided drug design techniques have accelerated the screening strategy in drug discovery. ¹¹ In silico docking experiment in the herbal drug candidate screening can accelerate the process, significantly reducing cost.

Sr. No	Botanical Name	Biological Source (B.S) & Family (F)	Phytoconstituents	Activity
1.	Antrodia cinnamomea	B.S:- Inner heartwood of the tree Cinnamomum Kanehira. F:- Fomitopsidaceae	Triterpenoids (17-18 %) , polyphenols (13-14%), Flavonoids (1-12%) Polysaccharides	Antiinflammatory, Anti-metastatic, Antioxidant, antiangiogenesis
2.	Ficus hirta	B.S :- The fruit and roots of the plant are usually referred to as hairy figs . F:-   Moraceae	flavonoids, coumarins, phenylpropanoids,  steroids, triterpenoids, phenolic acids.	Hepatoprotective, Anti-cancer,  Anti-inflammatory, rheumatism, hepatitis, and fatigue relief
3.	Terminalia catapa  linn	B.S:- Terminalia catapa Plant . F:- Combretaceae	Phenolic compounds , Essential oils	Anti-inflammatory, Anti-fungal, Antioxidant, Anticancer, Hypoglycaemic effects.
4.	CANNABIS SATIVA L.	B.S:- cannabis plant F:- Cannabaceae	2-carboxylic acids (2-COOH), 9-tetrahydrocannabinol cannabidiol (CBD), Cannabino	convulsions , seizures, anxiety,  nausea, antiinflammatory
5.	scutellaria baicalensis (Baikal skullcap)	B.S:- Baikal skullcap, also known as Scutellaria baicalensis, is a type of flowering plant. F:- Lamiaceae	Baicalin, glycoside baicalin, wagonin, wogonside	Antioxidant, Anti-inflammatory, Anti-allergic , Anti-tumorigenic, Anti-cancer
6.	Myricaceae	B.S:- This includes the parent genus Myrica, or sweet gales, which is a tiny family of dicotyledonous shrubs and small trees . F:- Myricaceae	Myricadiol, lasaxerol, lasaxesone, flavonoids , phenols	Anti-cancer, hepatoprotective, Antibacterial
7.	Lycopodium clavatum (Clubmoss)	B.S:- The biological source of Lycopodium clavatum is the plant itself, specifically the spores F:- Lycopodiaceae	Alkaloids (0.2-0.6%), Triterpenes and sterols (0.5-2%), Lipids and fats (1-3%), Polysaccharides (5-10%), Spores (60-80%).	Neuroprotective activity (Alzheimer's disease),  Liver protective activity, Anticancer activity.
8.	Diaryheptanoid Myricanone	B.S:- Myrica rubra , Myrica esculenta , Myrica gale . F :- Myricaceae	Myricanone  (0.5 – 2.5 %) , Myricanone A & B  (0.2 – 1.5 %), Myricano ( 0.8 – 3.0 %) Flavnoids –(2-5%) Triterpenoids 1.5-4.0 % polyphenols & tannins  3-7 % Essential oils 0.5-2.0.	Hepatoprotective, anticancer, Anti-inflammatory , antioxidant .
9.	CHELIDONIUM MAJUS ( Celandine)	B.S:- The bigger celandine, or Chelidonium majus, is a perennial herbaceous blooming plant. F:- Papaveraceae	Isoquinoline alkaloids, Flavonoids.	Anticolorectal cancer , Anti-pancreatic, Anti-cervical cancer activity, Biliary disorder management, Antimicrobial activity, Anthelmintic activity.
10.	Hedyotis Diffusa (Pusa F)	B.S:- Whole dried head of the plant Hedyotis diffusa Willd. F :- Rubiaceae	Iridoids, Flavonoids, Anthraquinones, Polysaccharides, Phenolics.	Anti-cancer , Anti-inflammatory,  Anti-oxidant, Hepatoprotective.
11.	Andrographis paniculata	B.S :- The entire plant, particularly the aerial parts. F :- Acanthaceae	Andrographolide,  Polyphenols,  Flavonoids,  Deoxyandrographolide,  Dilophenol,  Neoandrographolide	Anticancer, Anti-allergic, Antiviral, Immunostimulatory.
12.	Rhus coriaria	B.S:- The leaves of a flowering shrub. F:- Anacardiaceae	Hydrolysable tannins, Flavonoids, Anthocyanins, Phenolic acids	Anti-cancer,  Antioxidant,  Hypoglycemic,  Hypolipidemic
13.	Duchesnea indica	B.S:-perennial ground-hugging plant, also known as Indian Strawberry or Imitation Strawberry. F:- Rosaceae	Sterol, lignins,  tannins, terpenoids, and flavonoids	Induction of apoptosis, Anti-proliferation, inflammatory activities, anti-viral,  antimicrobial activities
14.	Crocus sativus L.	B.S:- The dried red stigmas of a perennial herb, commonly known as Saffron. F:- Iridaceae	proteins, amino acids,  minerals,  mucilage,  vitamins (particularly riboflavin and thiamine), lipophilic and hydrophilic carbohydrates,  colors such as crocin, anthocyanin, carotene,lycopene, zigzantin,  flavonoids,  starch, gums, and other chemical substances.	Induction of apoptosis,  Anti-proliferative, Antioxidant, Anti-inflammatory, Neuroprotective
15.	Trichosanthes kirilowii	B.S:-Trichosanthes kirilowii is a perennial climbing plant, native to Asia. Its fruit and root have been used in traditional Chinese medicine for various health purposes. Fruit (pericarp), Root   F:- Cucurbitaceae	terpenoids,  flavonoids,  alkaloids,  phytosterols, and lignans	Antitumor, Anti-inflammatory
16.	Moringa Oleifera	B.S :- Leaves of Moringa oleifera plants   F:- Moringaceae	9-octadecenoic acid (20.89%), L-(+)-ascorbic acid- 2, 6-dihexadecanoate(19.66%), 14–methyl-8-hexadecenal (8.11%), 4- hydroxyl-4-methyl-2-pentanone (7.01%), 3-ethyl-2.	Anti-inflammatory, neurodegenerative and cancer
16.	Origanum vulgare (Oregano)	B.S:- The plant , o. vulgare was been collected known as oregano .   F:- Lamiaceae	Cyclohexane, 1,5-diethenyl-2 ,3 dimethyl-, (1.alpha., 2.alpha., 3.alpha., 5.alpha.), Sesquiterpene epoxides, Fatty acid methyl esters,	malaria,  liver disease,  stress, inflammation, hypertension, influenza,  digestive problems,  kidney problems,  and skin, liver, stomach, lung, prostate, and colon cancer
17.	Cymbopogon citratus ( Lemongrass)	B.S:-Cymbopogon citratus is a perennial grass   F:- Poaceae  (Gramineae)	Cyclotrisiloxane, hexamethyl-, Propionic acid, Silane, diethylpentadecyloxy(2 phenylethoxy),	preservatives and pesticides, liver disease,  stress , malaria , inflammation ,  hypertension ,  influenza .
18.	Ganoderma applanatum	B.S:-Ganoderma applanatum is parasitic and saprophytic and grows as a mycelium within the wood of living and dead trees.   F:- Ganodermataceae (Basidiomycete)	alkaloids,  steroids, polyphenols, polysaccharides,  fatty acids,  micronutrients,  minerals, and vitamins, γ-terpinene, Dlimonene, cis-2- methyl-4-pentylthianes, s-dioxide, β-cymene, and α-terpinolene	Antioxidant, anticancer,  anti-inflammatory,  and immunomodulatory
19.	Boswellia serrata	B.S:- The resin, or gummy exudate, obtained from the bark of the tree   F:- Burseraceae	Terpenoids, Phenols, Polysaccharides	Anti-inflammatory,  Anti-microbial,  Anti-tumour, Anti-arthritic,  Anti-cancer.
20.	Portulaca oleracea	B.S:- Whole plant   F:- Portulacaceae	Flavonoids, fatty acids, alkaloids,  terpenoids, polysaccharides.	Anti-microbial,  anti-cancer, antidiabetic, anti-ulcerogenic.
21.	Bolbostemma paniculatum	B.S:- Dry tuber of plant.   F:- Cucurbitaceae	Sterols,  triterpenoids, alkaloids	Anti-tumor, anti-viral, anti-inflammatory.
22.	Agrimonia eupatoria ( “agrimony”)	B.S:- Whole plant   F:- Rosaceae	Steroids,  saponins, carbohydrates,  flavonoids, and phytoconstituents	Anti-inflammatory, neuroprotective,  Anti-diabetic, hepatoprotective,  and anti-cancer.
23.	Camellia sinensis (Green Tea)	B.S:- leaves of the shrub/small tree of the flowering plant.   F:- Theaceae	Flavonoids, Carotenoids, Tannins, Steroid.	Antioxidant,  Anti-inflammatory,  Anti-cancer
24.	Echinacea purpurea (Purple Coneflower)	B.S:- It obtained from roots, aerial parts, flowers. F:- Asteraceae	Alkaloid, Polysaccharides, Glycoproteins,  Caffeic acid	Immune system modulation, Anti-inflammatory, Antioxidant.

1.) Terminalia catappa

A study by Majoumouo et al. assessed the cytotoxic ability of “Terminalia catappa” endophytic fungal extracts against human cervical cancer cells, indicating their potential as promising anticancer agents. Terminalia catappa is a widely used traditional medicinal plant which has shown promising action in the treatment of cervical cancer by its endophytic fungi extract . The cytotoxicity of this extract was estimated in this study , with a particular attention to the N97 extract which is obtained from the stem bark. At half-maximal inhibitory concentration (IC50) of 33.35 µg/ml, the N97 extract have shown a strong anti-proliferative effects on cervical cancer  (HeLa) cells. The extract also demonstrated selective cytotoxicity, demonstrated by a cytotoxic concentration (CC50) of 268.4 µg/ml against human foreskin fibroblast (HFF) cells which were not malignant , thereby resulting in selective index of 8.01 . This shows that the extract can target cancer cells more specifically  without causing any damage to the healthy cells . The Annexin V/PI flow cytometry study conclusively demonstrated that the N97 extract induces apoptosis in HeLa cells.  Early and late apoptosis indicators significantly increased as a result of the apoptotic impact, indicating that it may have anticancer properties.  Terminalia catappa is a potential therapeutic and one of the natural treatment option for the treatment of cervical cancer . Further studies are authorized in order to isolate and identify the specific bioactive compounds which are responsible for these action .²¹

2.)  Lycopodine

The study by Mandal et al. showed the anti-cancer activity of “Lycopodine”, a bioactive chemical that is isolated from Lycopodium clavatum, in preventing HeLa cell proliferation by inducing cell death through capase 3 activation. Lycopodium clavatum's lycopodine's possible mode of action and therapeutic impact on cervical cancer. In this study, HeLa cells is a cervical cancer cell line, which are used to evaluate anti-cancer apoptotic effect of lycopodine. The chemical triggered caspase-3, which is responsible for programmed cell death. This caused the DNA in the cells to break down. Cell DNA fragmentation was indicated by increased sub G1 cell populations. Additionally, the study found that lycopodine administration resulted in a decrease in mitochondrial membrane potential and the release of cytochrome C into the cytoplasm. Additionally, the production of ROS increased, which triggered the apoptotic cascade. The substance also decreased the anti-apoptotic protein Bcl-2 and increased the pro-apoptotic protein Bax, suggesting that it helps in destroing cancer cells. Lycopodine is a promising chemotherapeutic cervical treatment drug for this malignancy.²²

3.) Antrodia cinnamomea

With a focus on their modes of action and the function of herbal formulations in cancer management, Ray and Pau investigate the therapeutic potential of Chinese herbs in the treatment of cervical cancer. “Antrodia cinnamomea” is a medicinal mushroom which is traditionally used in the Chinese medicinal system and has shown significant action in treating cervical cancer. The study shows that extract obtained from this fungus have anticancer effects by multiple mechanisms and primarily by inducing apoptosis in cancer cells .  Studies was been carried on human cervical cancer cells, such as HeLa and c-33a which shows that Antrodia cinnamomea triggers the apoptotic pathways by activating caspase enzyme and thereby increasing the cystolic levels of cytochrome c. It modulates the expression of key proteins, by elevating the pro-apoptotic markers like Bad , Bak, and Bim while downregulating anti-apoptotic proteins such as Bcl-2. Along with, apoptosis-causing properties, the extract has shown to inhibit the growth of cancer cells by causing cell cycle arrest, especially during the G2/M phase . According to these findings, ‘Antrodia cinnamomea’ shows a promise as an adjuvant therapy for cervical cancer complementing conventional treatments and potentially mitigating adverse effects To confirm its effectiveness and guarantee its safe incorporation into contemporary cancer therapy procedures, more clinical trials are necessary.²³

4.) Gonolobus condurango

Bishayee et al. examine the anticancer potential of “Gonolobus condurango” (Condurango) extract, showing that it can depolarize the mitochondrial membrane and activate the Fas receptor, causing ROS-dependent apoptosis in HeLa cells in vitro. Condurango extract (CE) , is derived from Gonolobus condurango and has shown high potential in the treatment of cervical cancer. Condurango extract causes programmed cell death through reactive oxygen species (ROS) according to research conducted on HeLa cells, a cervical cancer cell type. When HeLa cells are exposed to condurango extract, HeLa cells experience mitochondrial membrane depolarization, which leads to the release of cytochrome c and thereby leading to activation of caspase enzyme, subsequently leading to cell death. Additionally, condurango extract downregulated nuclear factors kappa-light-chain-enhancer of activated B cells (NF-κB) and B-cell lymphoma 2 (Bcl-2). Mitochondrial and receptor mediated apoptosis are involved in the dual pathway activation, which highlights the Condurangos potential as a therapeutic drug .²⁴

5.) Moringa oleifera

Indian researchers Pratibha Pandey and Fahad Khan present a unique therapeutic paradigm for the treatment of cervical cancer that centers on Jab1 inhibition. This study shows the anti-cancer properties of methanolic extract of leaves of “Moringa oleifera” (MMlE) aginst cervical cancer cells (HeLa). Soxhlet extractor was used in order to obtain MMLE extract, and is further tested for cytotoxicity via MTT assay. This showed a dose and time-dependent decrease in the viability of cells. Further investigations revealed that MMLE induced apoptosis, which was verified by NAC (ROS inhibitor), nuclear condensation (Hoechst 33342 staining), and elevated intracellular ROS levels. Additionally, MMLE affects gene expression by downregulating Jab1 and upregulating p27, which causes cell cycle arrest in the G0/G1 phase. Additionally, MMLA induced apoptosis was confirmed by flow cyclometric analysis, and its effects on important regulatory genes were confirmed by RT-PCR studies. The findings suggests that MMLE induces significant anti-cancer effects through apoptosis induction and cell cycle modulation .This study emphasis the therapeutic activity of MMLE in the cervical cancer treatment , warranting further in vivo and clinical investigation .²⁵

6.) Cymbopogon citrus (DC.) Stapf  (lemongrass) and origanum vulgare linn (oregano)

An insightful study by Aminat Omope Yusuf et al. explored the anticancer properties of the extract “Cymbopogon citrus  Stapf” and “origanum vulgare Linn”, in this study, use of  GC.MS analysis was done in order to identify bioactive compounds present in the extract along with its therapeutic potential. Cymbopogon citrus (lemongrass) and Origanum vulgare (oregano) extracts were studied for their potential to prevent cervical cancer. N-hexane, dichloromethane (DCM), and ethyl acetate were used to treat and fractionate the plants, which were gathered from Plateau State, Nigeria.  Different concentrations of extract were applied to the HeLa cervical cancer cells and cytotoxicity was been evaluated using the MTT assay and with the use of Graph Pad prism, CC50 values where determined. GC-MS analysis identified phytochemical compounds in the aqueous extracts where O. vulgare had a CC50 of 170.2 µg/mL, but C. citratus had 43.43 µg/mL.  By using vincristine as reference (0.75 µg/mL), the n-hexane fraction of C. citratus showed the strongest effect (CC50 = 3.81 µg/mL), followed by its DCM fraction (5.44 µg/mL). The strongest anti-cervical activity was exhibited by the n-hexane fraction of  origanum vulgare. This shows the potential of cybopogon citrus and origanum vulguare, specifically their n-hexane fractions, which are a high achieving treatments for cervical cancer.²⁶

7.)Portulaca oleracea

Khatibi S et al. conducted an in vitro study in order to evaluate the cytotoxic and antiproliferative effects of “Portulaca oleracea” ethanolic extract on HeLa cervical cancer cells. The study showed the antiproliferative effects of Portulaca oleracea ethanolic extract on HeLa cervical cancer cells. The extract was prepared by powdering the aerial parts of the plant and further percolating it in 80% of ethanol which is further followed by ultrasonic treatment, filtration and drying. Cell proliferation showed a significant decrease by the demonstration of the MTT assay in a dose-and time dependent manner, where the highest cytotoxicity was recorded to be  1500 µg/mL after 48 hours (P < 0.001) . These results were confirmed by the Tryptan Blue exclusion test, which showed reduced cell viability at concentrations of 700, 1000 , 1200 and 1500 µg/mL . Statistical analysis using ANOVA followed by Tukey’s test revealed significant differences between control groups (P < 0.05). To ascertain p. oleoracea extracts safety and effectiveness in preclinical models and normal cells, further studies are necessary to clarify the mechanisms, optimize dosages and evaluate its clinical application. Future research should be in such a way that it concentrates on confirming the animal models.²⁷

8.)Excoecaria agallocha L.

Tamanna Sultana et al. showed the anti-cancer potential of the leaves of “Excoecaria agallocha L.” extract, which shows action against the cervical cancer cells (SiHa). This study aimed at the assessment of the cytotoxic effects and the underlying the mechanism of action .The study shows that leaf extracts from Excoecaria agallocha L. have the potential to be targeted therapeutic agents, which are important in the treatment of cervical cancer. Silica gel column chromatography fractionation showed cytotoxicity against SiHa cervical cancer cells, with one fraction exhibiting an  IC50 of 15.538 ± 0.577 µg/mL. Normal peripheral blood mononuclear cells were not harmed. Bergenin is one of the major essential components in mediating the extract's anticancer activity . The formation of LC3 puncta in EGFP-SiHa cells provided evidence that the extract enhanced mitophagy and promoted autophagy. Furthermore, it triggered apoptosis by SMAC-induced cytochrome c release, as shown by caspase-3 activation. G2/M phase cell cycle arrest was demonstrated by Western blot analysis and flow cytometry, with lower levels of cyclin B1, cyclin D1, and Cdc2 and increased expression of p21 and p53.The combined admission of autophagy, apoptosis, mitophagy, and cell cycle arrest shows E. agallocha as a promising anti-cancer agent. This mechanism suppresses the cancer cell growth synergistically. This research paves the way for the development of plant-based treatments that fight against cervical cancer without endangering healthy cells .²⁸

9.)Imperata cylindrica

To find possible bioactive components that may be responsible for the therapeutic effects of “Imperata cylindrica” root extract, Paul Nayim et al. examine the root extract's in vitro anticancer activity against human cervical cancer cells. This study shows the Anti-cancer potential of Imperata cylindrica root (IC-MeOH) extract which is used in the management of cervical cancer. The extract establishes dose- and time-dependent cytotoxic effects when evaluated on HeLa and CaSki cervical cancer cells . . IC50 values were lower for CaSki cells, indicating higher sensitivity compared to HeLa cells . The extract lead to inhibition of cancer cell proliferation , followed by reduced colony formation and  apoptotic cell death . G0/G1 cell cycle arrest was confirmed by flow cytometry analysis, which stopped the proliferation of further cancer cells. However, IC-MeOH was less toxic to normal cervical cells (HCK1T) and demonstrated potential cytotoxicity when compared to cisplatin. Bioactive compounds maybe responsible for extracts anti-cancer effects by the use of UHPLC-HRMS analysis . This study shos that Imperata cylindrica might be a natural theraphy for the cervical cancer treatment  , but it is necessary to carry out further studies in order to confirm its efficacy . ²⁹

10.) Indigofera aspalathoides (Vahl.)

"Venkatesan Ramya et al. conducted a study on the cytotoxic activity of “Indigofera aspalathoides (Vahl.)” extracts in cervical cancer (HeLa) cells and demonstrated that ascorbic acid adjuvant treatment enhances the cytotoxic effects". Determining the cytotoxic potential of L. aspalathoides and clarifying its ethnopharmacological application as an adjuvant in lime juice (ascorbic acid) are the goals of this study.  Following the extraction of the active chemicals identified in I. aspalathoides using the Soxhlet extraction process with methanol and ethanol as solvents, and analytical assessnent was performed using gas chromatography combined with mass spectrometry (GC-MS). By examining the DPPH radical scavenging activity, ferric reducing antioxidant capacity (FRAC), and ABTS cation radical scavenging activity, we evaluated the in vitro antioxidant efficiency. The cytotoxic effects of SV-ME and EE against normal murine fibroblast (L929) cells and human cervical cancer (HeLa) were assessed using the MTT test. Hoechst dye and a dual acridine orange/ethidium bromide (AO/EB) fluorescent probe were used to assess the cytotoxicity and apoptosis induction. The mitochondrial membrane potential (Δψm) was measured using JC1.  In order to examine the additive and synergistic effects of ascorbic acid (AA), HeLa cells were co-treated with AA at IC doses in addition to SV-ME and SV-EE. Higher DPPH radical scavenging activity was demonstrsted at 200 μ 50 g/mL by SV-ME and EE.  Both extracts showed good ferrous reduction antioxidant capacity and ABTS radical cation scavenging action.  Furthermore, SV-ME and SV-EE treatment increased nuclear condensation and DNA fragmentation in normal (L929) cells in a dose-dependent manner while decreasing cell viability and causing apoptosis in malignant (HeLa) cells . Both extracts enhanced the depolarization of the mitochondrial transmembrane potential (Δψm) and modify the cellular redox status in HeLa cell lines. Additionally, increase in cytotoxic activity of SV-ME and SV-EE in HeLa cells was observed with ascorbic acid (AA) therapy. I. aspalathoides applied in Siddha medicine which has an potential cytotoxic effect against cervical cancer cells (HeLa) .³⁰

11.) Abrus precatorius

Kaur et al. showed the antiproliferative efficacy of “Abrus precatorius” seed extracts on cervical carcinoma by using invitro techniques and thereby highlighting its potential as a natural therapeutic agent. The seeds of the medicinal plant Abrus precatorius are shown to have promising therapeutic effects. In this study the antioxidative and antiproliferative qualities of the seed extract (ethyl acetate and 70% ethanol) where produced by using Soxhlet extraction technique which were further evaluated in this study against Hep2c and HeLa cells . APA (Mac) extract  was seen to have high phenolic content , APE (Sox) extract had a greater level of total flavonoids, and the APA (Sox) extract had a high level of total tannins. With this , free radical scavenging activity was demonstrated by APA (Sox) inorder to determine the maximum activity . APE (Mac) was the most effective inhibitor of Hep2C cells. APA (Sox) proved to be the most potent inhibitor of HeLa cells growth. whereas APE (Mac) extract had the most SOD, catalase, and GSH. APA (Mac) extract had the ultimate GST activity in Hep2C cells. It also had the lowest MDA level. The APA(Sox) extract has the maximum SOD, GST,GSH and the lowest MDA, while the (Mac) extract as the best catalase activity in HeLa cells. According to molecular docking studies, tannic acid binds most strongly to the GCR and HER2 receptor. This study demonstrates that A. precatorius seed extracts has intresting bioactive molecules that might fight aganist cervical cancer and safeguard cell from injury.³¹

12.) Berberis aquifolium

Singh et al. investigated “Berberis aquifolium's” potential to target aberrant expression of HPV oncoproteins, STAT3 oncogenic transcription factors, and AP-1 cancer promoting transcription factors in cervical carcinoma, providing insight into its potential therapeutic applications. The study investigates the accessibility of anti-cervical cancer (CaCx) and anti-HPV characteristics in phytochemical preparations made from the root of the berberine plant, B. aquifolium, with the goal to ease the repurposing of the medicine based on B. aquifolium in the treatment of CaCx. The study's primary goal was to assess the molecular oncogenic potential of various doses were B. Aqui folium root mother tincture (BAMT) ethanolic extract damaged the carcinogenic potential of HPV-positive (HPV16:Siha, HPV18:HeLa) and HPV-negative (C33a) CaCx cell lines. The MTT assay was used to screen BAMT for anti-proliferative properties. Using flow cytometry, the course of the cell cycle was examined. The expression levels of STAT3, AP-1,HPV E6, and E7 was consider as immunoblotting, and nuclear localization was observed through fluorescence microscopy. "In silico molecular docking was employed to assess the potential of phytochemicals purportedly present in BAMT to inhibit HPV16 E6." BAMT reduced CaCx cell viability in a dose-dependent manner across all analysed cell types. In the G1 phase, cells treated with BAMT exhibit a sight but noticeable cell growth termination, according to flowcytometric analysis. BAMT therapy led to decreased protein expression of important transcription factors, including AP-1 complex components JunB and c-Jun, along with STAT3, showed a reduction in the activated form pSTAT3 (Y705). By reducing overall transcription factor activity, BAMT decreased the amount of transcription factors available in the cancer cells, but it did not stop remaining active transcription factors from entering the nucleus, according to immunocytochemistry. HPV E6 and E7 protein gradually decreased in BAMT-treated HPV-positive cells in tandem with these alterations. The strongest E6 inhibitory potential was demonstrated by ine. Molecular docking of active phytochemicals in B. aquifolium root suggested that the phytochemicals may impede with HPV-16 E6 associated with its critical cellular partners, p53 and E6AP. The compounds with the strongest E6 inhibitory potential were magnoflorine, palmatine, and berberine. The investigation illuminate the molecular mechanism of BAMT'S activity and testing of therapeutic potential against HPV infection and cervical cancer. ³²

13.) Vitex trifolia L.

The potentential therapeutic effect of rotundifuran, a labdane-type diterpene natural compound isolated from "Vitex trifolia L.", was indicated by gong et al.'s finding that Cyr61 is a potential target of inducing apoptosis in cervical cancer cells. The review examined the anti cervical cancer properties and probable causes of rotundifuran (RTF), is a natural chemicals derived from vitex trifolia l. We invented that RTF may stop cervical cancer cell lines, similarly HeLa and SiHa, by inducing apoptosis in vitro (IC50 less than 10 μM). The HeLa cell-inoculated xenograft model is used to further validate RTF's anticancer effects.  Additionally, the results showed that ROS-induced mitochondrial-dependent apoptosis through the MAPK and PI3K/Akt signal pathways may be linked to RTF's anticancer effects. Proteomics studies and drug affinity responsive target stability (DARTS) coupled mass spectrometry (DARTS-MS) indicated that Cyr61 may be an RTF target in cervical cancer cells. Future research on RTF as a potential treatment for cervical cancer might benefit from this work.³³

14.) Citrus hystrix, Citrus limon, Citrus pyriformis, and Citrus macrocarpa

A thorough investigation into the phytochemical makeup, antioxidant qualities, and antiproliferative effects of essential oils extracted from leaves of “Citrus hystrix”, “Citrus limon”, “Citrus pyriformis”, and “Citrus macrocarpa”. The leaves distrub human cervical cancer cell lines was exhibited by Haneen Ibrahim, AI othman et al. This study evaluated the volatile components of essential oil produced from the leaves of C. hystrix, C. limon, C. pyriformis, and C.microcarpa from gas chromatograpy-quadeupole mass spectrometry. By using 84.88-97.99% of total ions and 80 secondary chemicals were tentatively identified. These comprised monoterpene hydrocarbons (5.20–76.15%), sesquiterpene hydrocarbons (0.21–38.87%), and oxygenated monoterpenes (3.91–89.52%). All citrus species included 27 different compounds, ranging in concentration from 1.19 to 39.06 percent. The loading parameters that affected these metabolic differences were also determined. The leading variables that contributed to these metabolic differences were also determined.Using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 2,2-diphenyl-1-picrylhydrazylhydrate (DPPH) tests, the antioxidant and antiproliferative properties of the citrus leaf oils were evaluated. With an IC50 value of 29.14 ± 1.97 mg/mL, C. limon had the strongest DPPH radical scavenging ability, meanwhile C. hystrix showed the lowest activity (IC50 value of 279.03 ± 10.37 mg/mL), as reported by the data. However, whole citrus oils have cell proliferation inhinition effects on the HeLa cervical cancer cell line; their IC50 values are 11.66 μg/mL for C. limon, 20.41 μg/mL for C. microcarpa, 25.91 μg/mL for C. hystrix, and 87.17 μg/mL for C. pyriformis.³⁴

15.) Peronema canescens  

“Peronema canescens” leaf extracts' anticancer properties against human cancer cell lines of HT-29, HeLa are investigated in vitro by Ibrahim Arsyik et al. To yet, the paper has only examined the bioactivity of Sungkai leaves (Peronema canescens Jack) in seawater shrimp larvae; no studies of the leaf's in vitro cytotoxic activity utilizing human colon and cervical cancer cells have been published.  This plant has a large number of secondary metabolites in its leaves that may have cytotoxic effects.  Herbal medications are natural therapeutic substances derived from plants that are used to cure cancer.  In vitro cell line assays can be used to test for a natural substance's anticancer efficacy. This work sought to ascertain the cytotoxicity (IC50) of P. canescens leaf extracts in ethanol, ethyl acetate, and chloroform against HeLa cervical cancer cells and HT-29 colon cancer cells.  Using a methanol solvent, the P. canescens leaf was extracted using the maceration process.  The liquid-liquid technique and consecutive polarity gradient eluents—ethanol, ethyl acetate, and chloroform—were used to separate the dried sample.  HT-29 and HeLa cells were utilized as cell lines for the MTT (3-(4,5-dimethylazole-2-yl)-2,5-diphenyltetrazolium bromide) cytotoxicity test.  A concentration range of 1.5 μg/mL to 200.0 μg/mL was employed.  Chloroform (10.353 μg/mL and 38.913 μg/mL), ethyl acetate (48.635 μg/mL and 28.186 μg/mL), and ethanol (42.017 μg/mL) had the highest cytotoxic activity values (IC50) on HT-29 and HeLa cells, respectively . The findings demonstrated that chloroform extracts' cytotoxic activity had a moderate impact on HeLa cervical cancer cells and a strong cytotoxic effect.  Opportunities for more research on anticancer are abundant.³⁵

CONCLUSION:

Cervical cancer remains a significant public health problem, especially in developing countries. While HPV vaccines have significantly reduced the onset of infection, therapeutic options for HPV induced cancers are minimal and are usually accompanied by several side effects. This review highlights recent advancements in drug discovery, including the identification and screening of plant-based phytoconstituents as potential anticancer agents. A number of phytoconstituents have shown promising results by targeting the HPV oncoproteins, restoring tumour suppressor functions, and modulating key signalling pathways involved in cancer progression. Plants like “terminalia catappa, moringa olifera, curcuma longa, and berberis aquifolium, among have shown effects by inducing apoptosis, regulating cell cycle, and exhibiting selective cytotoxicity against cervical cancer cells. These findings strengthen the therapeutic relevance of ethnomedical plants and prove the role of integrative approaches by the combination of traditional knowledge with modern drug discovery techniques. Future research should focus on in vivo validation, clinical trials, and the development of standardized herbal formulations for safe and effective cervical cancer therapy.

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