反义p55PIK慢病毒载体的构建及其对甲状腺癌细胞FTC236增殖的影响
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国家973计划 (2009CB521802);国家自然科学基金(30872472,30973496,30800569);湖北省自然科学基金(2008CDB174,2009CDB239);中央高校基本科研业务费专项资金(2011JC062,2011JC063);华中科技大学自主创新基金(2010MS027)


The antisense of p55PIK lentivirus could inhibit the proliferation of thyroid carcinoma FTC236 cells
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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-

    Abstract:

    Objective:To elucidate the role of antisense oligodeoxynucleotide of p55PIK inhibiting the proliferation of thyroid carcinoma cells. Methods:The lentivirus package system carrying the antisense RNA of p55PIK was constructed. The FTC236 cell was infected by the virus. Western blot and Realtime PCR were used to confirm the protein content and mRNA level of p55PIK. Then MTT assay and BrdU/PI corporation assay were used to analyze the proliferation of stable cells. Results:Two stable cell lines,one expressing p55PIK antisense RNA and the other expressing scramble antisense RNA,were constructed,and the p55PIK antisense RNA could knockdown the target protein’s expression(P < 0.01). In MTT assay,the absorbance ratio of the cells expressing p55PIK antisense RNA is (59 ± 19)% of that in the cells expressing scramble antisense. The BrdU/PI corporation assay showed that the DNA synthetize ratio of cells expressing p55PIK antisense RNA decreased by 11.6% compared with that of cells expressing scramble antisense. Conclusion:The p55PIK antisense can inhibit proliferation of the thyroid carcinoma cells by knockdown the p55PIK protein expression.

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李 川,金 源,来森艳,胡俊波.反义p55PIK慢病毒载体的构建及其对甲状腺癌细胞FTC236增殖的影响[J].南京医科大学学报(自然科学版),2012,(6):790-794

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  • 收稿日期:2012-01-07
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