基因沉默整合素连接酶对胰腺癌细胞上皮向间质转化的影响

来源 :中国实用医药 | 被引量 : 0次 | 上传用户:kunming
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目的探讨基因沉默整合素连接酶(ILK)对胰腺癌细胞(Panc-1)上皮向间质转化(EMT)的影响。方法培养Panc-1细胞,构建ILK-specific-sh RNA慢病毒载体后,再行转染Panc-1细胞,通过Western-blot技术验证感染基因的有效性,同时比较Panc-1细胞上皮向EMT转化标志性蛋白E-钙连蛋白(E-cadherin)表达情况。结果 (1)基因沉默ILK明显的抑制了Panc-1细胞的迁移、侵袭能力,且阳性组细胞迁移率和侵袭率均为(0.15±0.03),明显低于阴性组(0.43±0.02),差异具有统计学意义(t=23.572,P<0.01)。(2)SDS-聚丙烯酰胺凝胶电脉法(SDS-PAGE)结果 :E-cadherin的条带为122~141 KD,GAPDH为37 KD的条带。siR NA慢病毒转染Panc-1细胞后KD组E-cadherin表达为(0.545±0.011),明显高于阴性组(0.079±0.004),差异有统计学意义(t=28.942,P<0.01)。结论基因沉默ILK转染后,明显抑制Panc-1细胞迁移、侵袭能力,显著上调E-cadherin表达,逆转了EMT的发生。 Objective To investigate the effect of gene silencing integrin ligase (ILK) on the epithelial-mesenchymal transition (EMT) in pancreatic cancer cells (Panc-1). Methods Panc-1 cells were cultured and transfected into Panc-1 cells with ILK-specific-sh RNA lentiviral vector. The efficiency of infection gene was verified by Western-blot. Meanwhile, the transformation of Panc-1 cells into EMT The expression of landmark protein E-cadherin. Results (1) Gene silencing ILK significantly inhibited the migration and invasion ability of Panc-1 cells, and the positive cell migration rate and invasion rate were (0.15 ± 0.03), which were significantly lower than those in the negative group (0.43 ± 0.02) Statistically significant (t = 23.572, P <0.01). (2) Results of SDS-polyacrylamide gel electrophoresis (SDS-PAGE): The band of E-cadherin was 122-141 KD and the band of GAPDH was 37 KD. The expression of E-cadherin in KD group after transfected with siR NA lentivirus was (0.545 ± 0.011), significantly higher than that in the negative group (0.079 ± 0.004), the difference was statistically significant (t = 28.942, P <0.01). Conclusion The gene silencing ILK transfection significantly inhibits the migration and invasion of Panc-1 cells, up-regulates the expression of E-cadherin and reverses the occurrence of EMT.
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