Characterization and apoptotic effect of copper nanoparticles biosynthesized fromZiziphus zizyphusleaf on human renal cell carcinoma A498 cells

Biological formulation of copper nanoparticles (Cu-Nps) in cancer treatment has augmented due to its beneficial and curative effects. Ziziphus zizyphus(ZZ) is a species of medicinal plant traditionally used for various ailments. Hence, this work deals with formulation of copper nanoparticles (Cu-NP's) fromZ. zizyphusleaf extracts (Cu-ZZ NP's) and examine its effect on human renal carcinoma A498 cells. The characterization of the formulated Cu-ZZ NPs was done by spectroscopic and microscopic methods. The anti-cancer tests (MTT, ROS, MMP, Hoechst staining, comet, Real-time PCR, and western blot) were used to evaluate the biological properties of Cu-ZZ NPs on the growth and proliferation of A498 cells. The cytotoxic activity of Cu-ZZ NPs resulted in potent toxicity of A498 cells. Further, Cu-ZZ NP's triggered an intracellular ROS generation through mitochondrial pathways in the A498 cells, revealing a noteworthy cytotoxicity activity. Also, fluorescent microscopic studies using Hoechst staining, and comet analysis confirmed the occurrence of apoptosis in Cu-ZZ NPs treated A498 cells. Following, quantitative RT-PCR presented a negative regulation ofBcl-2expression and positive regulation ofBid,Bax, and caspases (3 and 9) gene expression in Cu-ZZ NP's treated A498 cells. To further justify, inhibition of mTOR/PI3K/AKT protein expression by Cu-ZZ NPs in A498 cells, lead to an inference that Cu-ZZ NPs induce apoptosis in A498 cells by inhibiting the mTOR, PI3K/Akt pathway.

Automated summary

The study successfully prepared copper nanoparticles from Ziziphus zizyphus leaf extracts and demonstrated their cytotoxic effects on human renal carcinoma A498 cells. The Cu-ZZ NPs induced ROS generation and apoptosis gene expression, leading to significant cytotoxicity in the A498 cells. Additionally, the inhibition of mTOR/PI3K/AKT pathway by Cu-ZZ NPs was identified as a mechanism for inducing apoptosis in A498 cells.