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目的 :探讨稳定干扰整合素β3(integrinβ3,ITGB3)基因对人乳腺癌BT549细胞增殖的影响。方法:采用实时荧光定量PCR法和蛋白质印迹法检测乳腺癌BT549、MCF-7和MDA-MB-453细胞中ITGB3 m RNA和蛋白的表达。将干扰ITGB3表达的LV-ITGB3-sh RNA慢病毒感染BT549细胞后,应用实时荧光定量PCR法和蛋白质印迹法检测BT549细胞中ITGB3 m RNA和蛋白的表达水平,MTT法和FCM法检测BT549细胞的增殖能力和细胞周期,蛋白质印迹法检测BT549细胞中c-Myc和cyclin D1蛋白的表达情况。结果:乳腺癌BT549细胞中ITGB3 m RNA和蛋白的表达水平高于MCF-7和MDA-MB-453细胞(P值均<0.05)。LV-ITGB3-sh RNA慢病毒感染后,BT549细胞中ITGB3 m RNA和蛋白的表达水平低于阴性对照组(LV-NCsh RNA感染BT549细胞)和空白对照组(BT549细胞未进行感染)(P值均<0.01),细胞增殖能力增强(P<0.01),S期细胞所占百分比上升(P<0.01),c-Myc和cyclin D1蛋白的表达水平上调(P值均<0.05)。结论:稳定干扰BT549细胞中ITGB3表达后,可能通过上调c-Myc和cyclin D1蛋白的表达而促进细胞增殖。
Objective: To investigate the effects of stable interfering integrinβ3 (ITGB3) gene on the proliferation of human breast cancer BT549 cells. Methods: The expressions of ITGB3 mRNA and protein in breast cancer cell lines BT549, MCF-7 and MDA-MB-453 were detected by real-time fluorescence quantitative PCR and Western blotting. After BT-549 cells were infected with LV-ITGB3-sh RNA lentivirus that interferes with ITGB3 expression, the expression of ITGB3 mRNA and protein in BT549 cells was detected by real-time fluorescence quantitative PCR and Western blotting. MTT and FCM were used to detect the expression of Proliferation and cell cycle, Western blotting was used to detect the expression of c-Myc and cyclin D1 in BT549 cells. Results: The expression of ITGB3 mRNA and protein in breast cancer BT549 cells was higher than that in MCF-7 and MDA-MB-453 cells (all P <0.05). The expression of ITGB3 mRNA and protein in BT549 cells was lower than that in the negative control group (LV-NCsh RNA-infected BT549 cells) and the blank control group (BT549 cells were not infected) after LV-ITGB3-sh RNA lentivirus infection (P value (P <0.01). The percentage of cells in S phase increased (P <0.01) and the expression of c-Myc and cyclin D1 increased (P <0.05). Conclusion: Stably interfering with ITGB3 expression in BT549 cells may promote cell proliferation by up-regulating c-Myc and cyclin D1 protein expression.