川芎嗪对肾透明细胞癌增殖和侵袭作用及其机制的研究

Inhibitory effect of ligustrazine on renal clear cell carcinoma and its mechanism

目的:通过多种实验途径探讨川芎嗪对肾透明细胞癌A498细胞增殖和侵袭作用及作用机制,为临床防治肾透明细胞癌提供可行的治疗途径和方法。方法:以A498细胞为研究对象,在细胞水平采用MTT和平板克隆形成实验检测细胞川芎嗪对细胞增殖能力的影响;采用Transwell小室实验研究川芎嗪对肾透明细胞癌A498细胞的迁移和侵袭能力的影响;采用皮下接种和尾静脉注射的方法分别建立裸鼠皮下荷瘤模型和裸鼠体内转移模型,研究川芎嗪在裸鼠体内对肾透明细胞癌的增殖和侵袭作用的影响;通过Western blot检测研究川芎嗪是否抑制肾透明细胞癌上皮间充质转化(EMT),分别检测相对的蛋白分子。结果:MTT和平板克隆形成实验显示川芎嗪抑制了肾透明细胞癌A498的增殖能力;Transwell小室实验结果显示川芎嗪有效抑制了肾透明细胞癌A498细胞的迁移和侵袭能力;裸鼠皮下荷瘤模型结果显示,川芎嗪可以抑制肾透明细胞癌在裸鼠体内的增殖能力,同时也抑制了肾透明细胞癌在裸鼠体内迁移和侵袭的能力;Western blot结果显示不同浓度川芎嗪处理不同时间后的A498细胞中N-cadherin、Snail和Slug表达降低,差异具有统计学意义。结论:川芎嗪通过抑制肾透明细胞癌EMT机制从而抑制肾癌细的增殖、迁移和侵袭等恶性生物学行为。

Objective:To investigate the inhibitory effect and mechanism of ligustrazine on renal clear cell carcinoma A498 through various experimental approaches,and provide a feasible therapeutic approach and method for clinical prevention and treatment of renal cell carcinoma.Methods: A498 cells were used as the research object.The effects of tetramethylpyrazine on cell proliferation were detected by MTT and plate clone formation at the cellular level.Transwell chamber assay was used to study the migration and invasion of tetramethylpyrazine on renal clear cell carcinoma A498.The effects of subcutaneous vaccination and tail vein injection on the subcutaneous tumor-bearing model of nude mice and the metastasis model in nude mice were studied to study the effects of ligustrazine on the proliferation and invasion of renal clear cell carcinoma in nude mice.To observe the effect of ligustrazine on the morphology of renal clear cell carcinoma;qRT-PCR and WB were used to detect whether ligustrazine inhibited EMT of renal clear cell carcinoma and detected relative protein molecules.Results: MTT and plate clone formation experiments showed that ligustrazine inhibited the proliferation of renal clear cell carcinoma A498.Transwell chamber experiments showed that ligustrazine effectively inhibited the migration and invasion of renal clear cell carcinoma A498;The results of the tumor model showed that ligustrazine inhibited the proliferation of renal clear cell carcinoma in nude mice,and also inhibited the ability of renal clear cell carcinoma to migrate and invade in nude mice.The results of Western blot showed that the expression of N-cadherin,Snail and Slug decreased in A498 cells treated with different concentrations of ligustrazine for different time.The difference was statistically significant.Conclusion: Ligustrazine inhibits the malignant biological behaviors such as fine proliferation,migration and invasion of renal cell carcinoma by inhibiting the EMT mechanism of renal clear cell carcinoma.