Notch1基因表达的变化对人肾细胞癌ACHN细胞凋亡的影响

Effect of Notch1 gene expression on apoptosis of ACHN cells in human renal cell carcinoma

目的探究Notch1基因表达的变化和人肾细胞癌(RCC)ACHN细胞凋亡的关系。方法选取人RCC细胞株ACHN细胞为研究对象,采用不同浓度的姜黄素作用于ACHN细胞,收集不同时间段的ACHN细胞,采用实时荧光定量多聚酶链反应法(RT-qPCR).Western blot方法分析Notch1基因mRNA及蛋白的表达量变化。构建Notch1基因的ACHN细胞系,并在相同浓度姜黄素诱导相同情况下,采用流式细胞仪比较检测两种细胞的凋亡率。结果利用RT-qPCR检测不同浓度姜黄素诱导凋亡的ACHN细胞中Notch1基因的表达量,结果显示,Notch1基因随着姜黄素浓度的升高和作用时间的延长,其表达量随之升高(P<0.05);利用Western Bolt法检测不同浓度姜黄素诱导凋亡的ACHN细胞中Notch1蛋白的表达量,结果显示,Notch1蛋白随着姜黄素浓度的升高和作用时间的延长,其表达量也随之升高(P<0.05)。姜黄素诱导48 h后,ACHN细胞(Notch1+)和敲除Notch1基因的ACHN细胞(Notch1-)的凋亡率分别是(4.902±0.163)%和(9.097±0.602)%,两者差异有统计学意义(P<0.05)。结论Notch1能够促进ACHN细胞凋亡,可为RCC的研究提供更多的理论依据,也为RCC的临床治疗提供新的思路和方法。

Objective To explore the effect of Notch1 gene expression on apoptosis of ACHN cells in human renal cell carcinoma(RCC).Methods The human RCC cell line ACHN cells were used as the research object.The ACHN cells were treated with different concentrations of curcumin.The ACHN cells were collected at different times,and then the changes of Notch1 gene mRNA and protein expression were analyzed by fluorescence quantitative Real-time PCR and Western blot.The ACHN cell line of Notch1 gene was constructed,and under the same condition that curcumin induced at the same concentration,the apoptosis rate of the two cells was detected by flow cytometry.Results Real-time PCR was used to detect different expression levels of Notchl gene in ACHN cells with different concentrations of curcumin-induced apoptosis.The results showed that the expression level of Notchl gene increased with the increase of curcumin concentration and the prolongation of action time(P<0.05).Western blot assay was used to detect the expression of Notchl protein in ACHN cells induced by curcumin at different concentrations.The results showed that the expression of Notchl protein increased with the increase of curcumin concentration and the prolongation of the treatment time(P<0.05).After 48 hours of curcumin induction,the apoptosis rates of ACHN cells(Notch1+)and Notch1 knockout ACHN cells(Notch1-)were(4.902±0.163)%and(9.097±0.602)%,re-spectively,there was significantly difference(P<0.05).Conclusions Notch1 can promote the apoptosis of ACHN cells,which can provide more theoretical basis for the study of RCC and provide new ideas and methods for the clinical treatment of RCC.