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目的探讨肽基脯氨酰同分异构酶(peptidyl-prolylcis/trans isomerase,Pin1)基因对子宫颈癌SiHa细胞生物学性能的影响。方法实验设立4组:空白对照组(正常培养的SiHa细胞)、阴性对照组(SiHa细胞转染随机序列)和Pin1siRNA1组、Pin1siRNA2组(转染siRNA序列1、2),采用基因RNA干扰技术下调子宫颈癌SiHa细胞中Pin1的表达;qRT-PCR和Western blot法检测Pin1 mRNA和Pin1蛋白水平的变化。Transwell小室体外实验检测Pin1基因对SiHa细胞侵袭、迁移能力的影响;应用流式细胞术分析其对SiHa细胞周期及凋亡的影响。结果 SiHa细胞转染基因Pin1siRNA干扰SiHa细胞后Pin1mRNA和Pin1蛋白表达水平明显降低;Transwell小室体外实验表明,Pin1siRNA1、Pin1siRNA2组蛋白表达下调后细胞侵袭、迁移能力降低;流式细胞术结果提示,与空白对照组、阴性对照组相比,Pin1基因被干扰后细胞阻滞在G0/G期,细胞凋亡率明显增高(P<0.05)。结论降低Pin1表达能明显抑制宫颈癌细胞的侵袭、迁移能力。Pin1参与了SiHa细胞的凋亡过程,并且随Pin1表达量的高低来调控肿瘤细胞的周期和凋亡程度。
Objective To investigate the effect of Pin1 gene on the biological behavior of cervical cancer SiHa cells. Methods Four experimental groups were established: control group (normal cultured SiHa cells), negative control group (SiHa cells transfected random sequence) and Pin1siRNA1 group, Pin1siRNA2 group (transfected siRNA sequence1,2), gene RNAi technology was used to down-regulate The expression of Pin1 in SiHa cells was detected by qRT-PCR and Western blot. The effects of Pin1 gene on invasion and migration of SiHa cells were detected by Transwell chamber in vitro. The effects of Pin1 on the cell cycle and apoptosis of SiHa cells were analyzed by flow cytometry. Results The expression of Pin1 mRNA and Pin1 protein in SiHa cells transfected with Pin1 siRNA were significantly decreased. The results of Transwell chamber showed that the expression of Pin1 siRNA1 and Pin1 siRNA2 decreased after invasion and migration. The results of flow cytometry Compared with the control group and the negative control group, Pin1 gene was blocked in the G0 / G phase and the apoptosis rate was significantly increased (P <0.05). Conclusions Reducing Pin1 expression can significantly inhibit the invasion and migration of cervical cancer cells. Pin1 participates in the process of SiHa cell apoptosis, and with the Pin1 expression level to regulate tumor cell cycle and apoptosis.