基因干扰miR-105抑制非小细胞肺癌细胞的干细胞样特性、恶性增殖和侵袭

Gene interference miR-105 inhibits the stem cell-like characteristics, malignant proliferation and invasion of non-small cell lung cancer cells

目的探究基因干扰miR-105对非小细胞肺癌细胞的干细胞样特性、恶性增殖和侵袭的影响。方法 RT-PCR检测33例肺癌患者肺组织及癌旁组织中miR-105表达水平。转染miR-105 inhibitor至A549细胞,将细胞随机分为3组(Control、inhibitor-NC、miR-105 inhibitor组)进行后续实验。RT-PCR检测miR-105表达水平;成球实验检测细胞成球率、成球直径;Weslern blot检测SOX2、OCT4蛋白表达水平;流式分选检测CD133+细胞数;克隆形成实验检测细胞增殖;Transwell检测细胞侵袭;Weslern blot检测VEGF蛋白表达水平。结果癌旁组织中miR-105表达水平低于癌组织。与Control组相比较,miR-105 inhibitor组miR-105表达水平降低(P<0.05),细胞成球率、成球直径降低(P<0.05),SOX2、OCT4蛋白水平降低(P<0.05),CD133+细胞数降低(P<0.05),克隆形成率降低(P<0.05),侵袭细胞数降低(P<0.05),VEGF蛋白水平降低(P<0.05)。结论基因干扰miR-105可抑制非小细胞肺癌细胞的干细胞样特性、恶性增殖和侵袭。

Objective To investigate the effects of gene interference with miR-105 on stem cell-like characteristics, malignant proliferation and invasion of non-small cell lung cancer cells. Methods RT-PCR was used to detect the expression level of miR-105 in lung tissues and adjacent tissues of 33 patients with lung cancer. After transfection of miR-105 inhibitor into A549 cells, the cells were randomly divided into three groups: Control group, inhibitor-NC group and miR-105 inhibitor group for follow-up experiments. The expression level of miR-105 was detected by RT-PCR. The pellet formation rate and pellet formation diameter were measured by pellet formation experiment. Western blot was used to detect the expression levels of SOX2 and OCT4 proteins. The number of CD133 positive cells was determined by flow separation. Cell proliferation was detected by clone formation assay. Transwell was used to detect cell invasion;Western blot was used to detect VEGF protein expression level. Results The results showed that the expression level of miR-105 in paracancer tissues was lower than that in cancer tissues. Compared with the Control group, miR-105 inhibitor group of miR-105 expression level decreased(P<0.05), the cells into a ball rate, into a ball diameter decreased(P<0.05), SOX2, OCT4 protein levels(P<0.05)and CD133 positive cells were lower(P<0.05), clone formation rate was lower(P<0.05), invasive cells were lower(P<0.05), VEGF protein levels were lower(P<0.05).Conclusion Gene interference with miR-105 can well inhibit stem cell-like characteristics, malignant proliferation and invasion of non-small cell lung cancer cells.