MTT与HOECHST染色法检测膜联蛋白A7基因沉默对Hela细胞的影响
陈雪, 李欣, 高福禄, 张金库
中国临床解剖学杂志 ›› 2014, Vol. 32 ›› Issue (1) : 57-60.
MTT与HOECHST染色法检测膜联蛋白A7基因沉默对Hela细胞的影响
The effect of Annexin A7 knockdown on Hela cells by MTT and HOECHST stain
目的 探讨膜联蛋白A7（ANXA7）低表达对人宫颈癌细胞系Hela细胞增殖和凋亡的影响。 方法 (1)体外培养Hela细胞，应用免疫组化和Western blot鉴定细胞膜联蛋白A7的表达，Western blot鉴定siRNA干扰对膜联蛋白A7表达的抑制效果；(2) MTT实验检测siRNA抑制膜联蛋白A7的表达后对Hela细胞增殖情况的影响；(3)激光共聚焦检测siRNA抑制膜联蛋白A7的表达后对Hela细胞凋亡情况的影响。 结果 (1)免疫组化和Western blot结果均显示Hela细胞内有膜联蛋白A7的表达。siRNA干扰效率的检测显示：siRNA干扰组与阴性对照组、空白对照组比较膜联蛋白A7的表达明显降低，差异有统计学意义（P<0.05）；(2) MTT实验显示：siRNA干扰组与阴性对照组、空白对照组比较细胞增殖速度无显著性差异（P>0.05）；(3)激光共聚焦检测结果显示：与阴性对照组和空白对照组相比，siRNA干扰组细胞凋亡率显著增加，差异具有统计学意义（P<0.05）。 结论 膜联蛋白A7的低表达可促进人宫颈癌细胞系HeLa细胞的凋亡。
Objective To study whether the knockdown of Annexin A7 can affect apoptosis and proliferation of HeLa cells. Methods (1)Immunohistochemistry and Western blot were used to identify the expression of Annexin A7 in human cervical carcinoma cell line Hela and to identify the inhibitory efficiency of siRNA interference on the Annexin A7 expression. (2) MTT assay was conducted to detect the impact on the proliferation of HeLa cells after the expression of annexin A7 was inhibited by siRNA. (3) Laser confocal microscope was conducted to detect the impact on the apoptosis of HeLa cells after the expression of Annexin A7 was inhibited by siRNA. Results (1) Annexin A7 was expressed in Hela cells and the Annexin A7 expression of siRNA group was reduced significantly (P<0.05), compared with that of the negative control group and the control group. (2)The cell proliferation of ANXA7siRNA group showed no significant difference compared with that of the negative control group and the control group (P>0.05). (3) The cell apoptotic rate of ANXA7siRNA group was significantly increased (P<0.05) compared with that in the negative control group and the control group. Conclusion The knockdown of Annexin A7 promotes the apoptosis of Hela cells.
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