LINC01018过表达结肠癌细胞株的建立及其生物学特性

Establishment of LINC01018 overexpression colon cancer cell line and study on its biological charac-teristics

目的建立LINC01018稳定过表达的结肠癌细胞株,并研究对其生物学特性。方法实时荧光定量PCR(qRT-PCR)检测人结肠黏膜上皮细胞HCoEpiC及人结肠癌细胞HT-29中LINC01018表达情况。采用LINC01018过表达慢病毒感染HT-29细胞,筛选稳定过表达LINC01018的HT-29细胞株。分别采用CCK-8实验.Transwell实验检测LINC01018过表达对HT-29细胞增殖、侵袭及迁移能力的影响。利用Western blot方法检测LINC01018过表达对HT-29细胞中CDK6及MMP-2蛋白表达的影响。结果LINC01018在HT-29细胞中的表达明显低于HCoEpiC细胞。成功筛选获得了稳定过表达LINC01018的HT-29细胞株HT-29-L18。在HT-29细胞中过表达LINC01018后,能够显著抑制HT-29细胞的增殖、侵袭以及迁移能力;同时CDK6及MMP-2蛋白表达明显下降。结论LINC01018在结肠癌细胞中异常低表达,LINC01018表达上调能够抑制结肠癌细胞的增殖、侵袭以及迁移能力,其机制可能与CDK6、MMP-2有关。

Objective To establish a colon cancer cell line which overexpressing LINC01018 stab-ly and study its biological characteristics.Methods The expression of LINC01018 in HCoEpiC and HT-29 cells were detected by real time fluorescence quantitative polymerase chain reaction(qRT-PCR).HT-29 cells were infected with LINC01018 overexpression lentivirus to screen and establish HT-29 cell lines which overexpressing LINC01018 stably.The effect of LINC01018 on the proliferation,invasion and migration of HT-29 cells were detected by cell counting kit-8(CCK-8)assay and Transwell assay separately.The ex-pression of CDK6 and matrix metalloproteinase-2(MMP-2)in HT-29 cells was detected by W estern blot.Results The expression of LINC01018 in HT-29 cells was significantly lower than that in the human colon-ic epithelial cells(HCoEpiC).HT-29-L18 cell lines which overexpressing LINC01018 stably was screened successfully.Overexpression of LINC01018 significantly inhibited the cell proliferation,invasion and migra-tion,and reduced the protein expression of CDK6 and MMP-2 in HT-29 cells.Conclusions The expres-sion of LINC01018 was decreased abnormally in colon cancer cells.U p-regulation of LINC01018 expression can inhibit the proliferation,invasion and migration of colon cancer cells,which may be related to CDK6 and MMP-2.