Department of Urology, Affiliated Hospital of Guangdong Medical University, 57 Renmin Street South, 524001, Zhanjiang, Guangdong, China.
Laboratory of Urology, Affiliated Hospital of Guangdong Medical University, 57 Renmin Street South, 524001, Zhanjiang, Guangdong, China.
Mol Biol Rep. 2023 Jul;50(7):5697-5707. doi: 10.1007/s11033-023-08381-w. Epub 2023 May 22.
Berberine is a natural isoquinoline alkaloid that has been shown to have antitumor properties in a growing number of studies. However, its role in renal cell carcinoma remains unclear. This study investigates berberine's effect and mechanism in renal cell carcinoma.
The methyl-tetrazolium, colony formation, and lactate dehydrogenase assay were used to detect proliferation and cytotoxicity, respectively. Flow cytometry, caspase-Glo 3/7 assay, and adenosine triphosphate assay were used to detect apoptosis and the adenosine triphosphate levels. Wound healing and transwell assay were used to examine the migration ability of renal cell carcinoma cells. Besides, the level of reactive oxygen species (ROS) was explored using a DCFH-DA-based kit. Additionally, western blot and Immunofluorescence assay was taken to determine the levels of relative proteins.
In vitro, our findings indicated that the proliferation and migration of renal cell carcinoma cells treated with berberine in various concentrations were inhibited, while the level of ROS and apoptosis rate were increased. Furthermore, The results of western blot showed that the expression of Bax, Bad, Bak, Cyto c, Clv-Caspase 3, Clv-Caspase 9, E-cadherin, TIMP-1and γH2AX were up-regulated, while Bcl-2, N-cadherin, Vimentin, Snail, Rad51 and PCNA were down-regulated after treating with berberine with various concentration.
The result of this study revealed that berberine inhibits renal cell carcinoma progression via regulating ROS generation and inducing DNA break.
小檗碱是一种天然异喹啉生物碱,越来越多的研究表明其具有抗肿瘤特性。然而,它在肾细胞癌中的作用尚不清楚。本研究探讨了小檗碱在肾细胞癌中的作用和机制。
采用甲基噻唑基四唑(MTT)比色法、集落形成实验和乳酸脱氢酶(LDH)检测试剂盒分别检测细胞增殖和细胞毒性,流式细胞术、Caspase-Glo 3/7 检测试剂盒和三磷酸腺苷(ATP)检测试剂盒检测细胞凋亡和 ATP 水平,划痕愈合实验和 Transwell 实验检测肾癌细胞的迁移能力,采用 DCFH-DA 试剂盒检测活性氧(ROS)水平,Western blot 和免疫荧光实验检测相对蛋白水平。
体外实验结果表明,小檗碱在不同浓度下处理肾癌细胞,其增殖和迁移受到抑制,而 ROS 水平和凋亡率升高。此外,Western blot 结果显示,用不同浓度的小檗碱处理后,Bax、Bad、Bak、Cyto c、Clv-Caspase 3、Clv-Caspase 9、E-cadherin、TIMP-1 和γH2AX 的表达上调,而 Bcl-2、N-cadherin、Vimentin、Snail、Rad51 和 PCNA 的表达下调。
本研究结果表明,小檗碱通过调节 ROS 生成和诱导 DNA 断裂抑制肾细胞癌的进展。