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目的探讨不同剂量X线照射对人结直肠癌Caco2细胞系中microRNA-221(miR-221)和张力蛋白在10号染色体上同源缺失的磷酸酶(PTEN)表达的影响。方法常规培养Caco2细胞系并分为5组,分别给予不同剂量(0、2、4、6及8 Gy)的X射线照射,24 h后提取细胞总RNA和蛋白质,应用real-time RT-PCR方法检测Caco2细胞中miR-221和PTEN mRNA的表达水平,用Western-blot法检测PTEN蛋白的表达水平。结果 Caco2细胞系在不同剂量的X线照射下,其miR-221表达水平随着照射剂量的增加而增高,呈剂量依赖性(P<0.01);PTEN mRNA的表达水平无明显变化(P>0.05),但PTEN蛋白表达水平则随X线照射剂量的增加而逐渐降低,亦呈剂量依赖效应(P<0.01)。结论 X线照射剂量可影响结直肠癌Caco2细胞中miR-221和PTEN蛋白的表达。
Objective To investigate the effect of different doses of X-ray irradiation on the expression of microRNA-221 (miR-221) and homocysteine-deleted phosphatase (PTEN) on chromosome 10 in human colorectal cancer Caco2 cell line. Methods Caco2 cell lines were routinely cultured and divided into 5 groups. X-ray irradiation with different dosages (0, 2, 4, 6 and 8 Gy) was performed. Total RNA and protein were extracted 24 hours later. Methods The expression of miR-221 and PTEN mRNA in Caco2 cells was detected by Western-blot, and the expression of PTEN protein was detected by Western-blot. Results The expression level of miR-221 in Caco2 cell line increased with the increase of irradiation dosage in a dose-dependent manner (P <0.01), while the expression level of PTEN mRNA did not change significantly (P> 0.05 ). However, PTEN protein expression decreased with the increase of X-ray dose (P <0.01) in a dose-dependent manner. Conclusion X-ray irradiation can affect the expression of miR-221 and PTEN protein in colorectal cancer Caco2 cells.