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目的:探讨siRNA抑制TCAB1表达对鼻咽癌CNE-2细胞放射敏感性的影响及分子机制。方法:构建靶向抑制TCAB1的siRNA干扰质粒及阴性对照质粒,转染至CNE-2细胞,RT-PCR和Western-blot检测TCAB1表达;克隆形成实验检测细胞放射敏感性参数D0、SF2等;QPCR检测转染前后CNE-2细胞端粒的相对长度变化。结果:成功构建CNE-2/phTCAB1-siRNA干扰质粒,转染后RT-PCR和Western-blot分析表明TCAB1mRNA和蛋白表达均受到明显抑制,干扰组细胞D0和SF2值分别为2.490和0.460,明显低于空白对照组(D0和SF2值分别为3.186和0.607),P<0.01;QPCR检测端粒相对长度发现干扰组T/S值(0.19±0.01)均小于正常细胞(0.52±0.06)和转染阴性质粒细胞(0.42±0.01),P<0.05,而正常细胞组和转染阴性质粒细胞T/S值比较差异没有统计学意义(P>0.05)。结论:抑制TCAB1表达可以通过影响端粒长度增加鼻咽癌CNE-2细胞的放射敏感性,TCAB1有望成为一种新的肿瘤放射增敏靶点。
Objective: To investigate the effect of siRNA on the radiosensitivity of nasopharyngeal carcinoma CNE-2 cells induced by TCAB1 and its molecular mechanism. Methods: siRNA interfering plasmids and negative control plasmids targeting TCAB1 were constructed and transfected into CNE-2 cells. The expression of TCAB1 was detected by RT-PCR and Western-blot. The radiosensitivity parameters D0 and SF2 were detected by clonogenic assay. The relative length of telomere in CNE-2 cells before and after transfection was detected. Results: The CNE-2 / phTCAB1-siRNA interference plasmids were successfully constructed. The expression of TCAB1 mRNA and protein were significantly inhibited by RT-PCR and Western-blot analysis after transfection. The D0 and SF2 values of the interfering cells were 2.490 and 0.460, respectively, which were significantly lower (D0 and SF2 values were 3.186 and 0.607 respectively), P <0.01; T / S values of interference group (0.19 ± 0.01) were lower than that of normal cells (0.52 ± 0.06) and transfection (0.42 ± 0.01), P <0.05. There was no significant difference in T / S between normal cells and transfected negative plasmids (P> 0.05). CONCLUSION: Inhibition of TCAB1 expression can increase the radiosensitivity of CNE-2 cells by affecting telomere length. TCAB1 is expected to become a new target for radiosensitization of tumors.