Rhodium (Ⅱ) Complex With 2-benzoylpyridine,Induces Apoptosis And Cell Cycle Arrest And The Mechanism In HepG2 Cells

Objective: Hepatocellular carcinoma(HCC)is one of the most commonly malignant tumors,which has a serious impact on human health.At present,HCC is mainly treated with surgical excision,radiotherapy and chemotherapy or combined therapy.However,due to the drug resistance,side effects and poor prognosis of chemotherapy drugs,it is strongly necessary to develop new anti-tumor drugs.Since cisplatin was synthesized and then used in clinical as an antitumor drug,metal complexes began to arouse more and more people’s attention.2-benzoylpyridine rhodium(II)complex is a new metal complex,owning to the biological activity,but its anti-tumor effect and the mechanism are still unclear.The main mechanisms of the new metal complex against Hep G2 cells,including the inhibition proliferation,apoptosis and cell cycle arrest,are researched in our article.Methods: 1.MTT was used to screen the best liver cancer cell line which was more sensitive to 2-benzoylpyridine rhodium(II)complex.2.The viability of HepG2 treated with 2-benzoylpyridine rhodium(II)complex was investigated by MTTand colony-formation assay.3.The apoptosis and cell cycle arrest of Hep G2 treated with 2-benzoylpyridine rhodium(II)complex were detected by flow cytometry.4.The morphological changes of apoptosis in HepG2 were observed by Hoechst staining and JC-1 staining.5.The level of reactive oxygen species(ROS)was detected by ROS assay kit and whether there was ROS-dependent or not.6.Western Blotting was used to detect the expression of apoptotic relative factors and cell cycle blocked relative factors.Results: 1.After the liver cancer cell lines treated with 2-benzoylpyridine rhodium(II)complex,HepG2 cell line was the most sensitive cell lines and then was selected for further study.2.The MTT results showed that 2-benzoylpyridine rhodium(II)complex decreased cell viability of HepG2 in time-and concertation-dependent manners.3.After treated with 40μM 2-benzoylpyridine rhodium(II)complex for 48 h,HepG2 cell apoptosis was detected by flow cytometry,the result owning a significant difference(P<0.05),and the nuclear condensed was probed by Hoechst 33258 staining,and the reduce of mitochondrial membrane potential was investigated by JC-1 staining.Moreover,2-benzoylpyridine rhodium(II)complex increased the level of reactive oxygen species(ROS),but that 2-benzoylpyridine rhodium(II)complex induced apoptosis was ROS-independent.4.The mitochondrial pathway and death receptor pathway involved in HepG2 apoptosis induced by 2-benzoylpyridine rhodium(II)complex werestudied by Western Blotting,and Bcl-2 families were also involved in the process.Besides,HepG2 cell cycle was blocked by upregulation the expression of p21 and downregulation the expression of cyclin A1,cyclin D1,cyclin E1,CDK2,CDK6.Conclusion: 2-benzoylpyridine rhodium(II)complex is a potential new anti-HCC drug,which induces HepG2 cell death via the mitochondrial and death receptor pathways and arrest cell cycle in G1 phase.