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目的:观察反义p55PIK慢病毒对甲状腺癌细胞FTC236的增殖是否有抑制作用。方法:构建含p55PIK反义RNA的慢病毒载体并包装成慢病毒,使之感染FTC236细胞,得到稳定的细胞系后用Western blot方法检测p55PIK蛋白的表达情况,用Real-time PCR检测p55PIK的mRNA的表达情况。同时用四甲基偶氮唑盐(MTT)法及BrdU掺入法检测各组细胞的增殖情况。结果:构建的慢病毒感染甲状腺癌细胞FTC236的效率约为95%;Western blot证明反义p55PIK病毒可以使该细胞的p55PIK蛋白水平降低,mRNA水平降为对照组的(47±8)%(P<0.01)。MTT法检测发现反义p55PIK慢病毒感染后的FTC236细胞增殖明显受到抑制,增殖率为未处理组的(59±19)%(P<0.05)。且反义p55PIK慢病毒组的平均BrdU掺入率比未处理组降低了10.64%(P<0.02)。结论:反义p55PIK病毒可以通过抑制p55PIK蛋白的表达而抑制了FTC236细胞的增殖。
Objective: To observe whether antisense p55PIK lentivirus can inhibit the proliferation of thyroid cancer cell FTC236. METHODS: A lentiviral vector containing p55PIK antisense RNA was constructed and packaged into Lentiviral vector to infect FTC236 cells. After stable cell lines were obtained, the expression of p55PIK protein was detected by Western blot. The p55PIK mRNA was detected by Real-time PCR. The expression of the situation. At the same time, the proliferation of cells in each group was detected by MTT method and BrdU incorporation method. RESULTS: The efficiency of the constructed lentivirus infected thyroid cancer cell FTC236 was approximately 95%; Western blot demonstrated that the antisense p55PIK virus could reduce the p55PIK protein level of the cell, and the mRNA level was reduced to (47±8)% of the control group (P). <0.01). The proliferation of FTC236 cells after antisense p55PIK lentivirus infection was significantly inhibited by MTT assay, and the proliferation rate was (59±19)% in the untreated group (P<0.05). The mean BrdU incorporation rate of antisense p55PIK lentiviral group was decreased by 10.64% compared with untreated group (P<0.02). CONCLUSION: Antisense p55PIK virus can inhibit the proliferation of FTC236 cells by inhibiting the expression of p55PIK protein.