Naveen Kumar AD

In Vitro Anticancer Activities of Senna singueana Leaf Extracts against HeLa Cell Lines

The present demonstrated that SSL-CH extract selectively inhibits the growth of HeLa cell lines though activation of p53 and Bax genes medaied proapoptotic pathway signifying it is a potential alternative anticancer agent for cervical cancer. The selective cytotoxicity of diethyl ether, chloroform, ethyl acetate, and ethanolic leaf extracts of S. singueana were evaluated against cervical HeLa and normal mouse fibroblast L929 cell lines using MTT assay. At (IC50) concentration 20 µg/ml) of effective chloroform leaf extract of S. singueana (SSL-CH) was chosen to evaluate its nuclear apoptotic changes by using Acridine Orange/Etidium Bromide dual staining.  Subsequently, apoptotic induction potential of SSL-CH extract was assessed by RT-qPCR analysis of apoptotic genes p53 and Bax. Treatment SSL-CH extract selectively inhibited the dose depended proflretion of HeLa cell lines with IC50 of 20 µg/ml. AO/EB dual staining showed SSL-CH extract induced late apoptosis of 49.42% in compared to the control. RT-PCR analyses demonstrated that SSL-CH extract profoundly upregulated the proapoptotic genes p53 to12.85 (***p≤0.01), Bax to 11.61 (**p≤0.01), caspase-9 to 40.62 (***p≤0.01), and caspase-3 to 11.61 folds (**p≤0.01) respectively and down regulated antiapoptotic genes Bcl-2 to 0.9 (**p≤0.001) and survivin gene to 1 (***p≤0.001) at 2×IC50 (40 µg/mL) in comparison to control, confirming activation of the intrinsic apoptotic pathway. These findings demonstrated that SSL-CH extract could serve as a potential alternative therapeutic agent for cervical cancer treatment with affordable and fewer side effects comparison with cisplatin.

Phytochemical Analysis and Apoptosis-Inducing Anticancer Activity of Buddleja polystachia Leaf Extract against HeLa Cell Lines

Cervical cancer is the second leading cause of mortality in women globally, with its incidence increasing due to multidrug resistance (MDR) and adverse side effects associated with chemotherapeutic agents. Hence, this study was carried out to investigate the selective cytotoxicity and apoptotic induction potential of Buddleja polystachya leaf diethyl ether (BPL-DE) extract on cervical cancer HeLa cell lines. The selective cytotoxicity of leaf extracts of B. polystachya was assessed by the MTT assay. The apoptotic induction potential of BPL-DE extract at IC₅₀ (20 µg/mL) was assessed by Acridine Orange/Ethidium Bromide (AO/EB) dual staining. Furthermore, qRT-PCR analysis was performed to study mRNA gene expression of pro-apoptotic (Bax, p53) and anti-apoptotic genes (Bcl-2, survivin), as well as caspase-9 and caspase-3 gene expression levels. The BPL-DE extract showed the highest selective cytotoxic effect against HeLa cells, with an IC₅₀ of 20 µg/mL, resulting in a selective index of 25.31. AO/EB dual-staining analysis revealed that the BPL-DE extract at IC₅₀ (20 µg/mL) significantly induced late apoptosis (p≤0.001). qRT-PCR results showed that the mRNA expression levels of Bax and p53 increased by 3.96-fold (p≤0.01) and 5-fold (p≤0.001), respectively. In contrast, Bcl-2 and survivin mRNA expression levels were significantly downregulated by 1.02-fold (p≤0.01) and 1.13-fold (p≤0.001), respectively, at two-fold IC50 (40 µg/mL). The BPL-DE extract also upregulated the mRNA expression of caspase-9 and caspase-3 by 32.07-fold (p≤0.001) and 15.06-fold (p≤0.01), respectively, at two-fold IC50. The findings of this study demonstrate that the BPL-DE extract significantly induced the p53-mediated intrinsic apoptotic pathway in HeLa cell lines and provides a potential alternative therapeutic agent for cervical cancer treatment by minimizing damage to normal cells.