miR-197靶向调节CD82对结肠癌细胞侵袭转移的影响
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泸州市科技计划项目


Effect of miR-197 on invasion and migration of colon cancer cells by targeting CD82
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    【摘要】 目的 探讨下调microRNA-197 (miR-197)对结肠癌HCT116细胞侵袭转移能力的影响及其作用机制。方法 将对数生长期HCT116细胞随机分为miR-197反义寡核苷酸(inhibitor)组、无义序列阴性对照(NC)组、空白对照(Mock)组,采用脂质体介导转染法,将miR-197反义寡核苷酸(miR197 inhibitor)转染HCT116细胞。运用荧光定量PCR (qRT-PCR)检测miR-197的表达水平;通过生物信息学预测miR-197靶基因,应用Western blot法检测CD82蛋白的表达;采用transwell小室实验检测HCT116细胞的侵袭、迁移能力的改变。结果 miR-197反义寡核苷酸转染HCT116细胞后,inhibitor组miR-197的表达较NC组和Mock组明显降低(P<0.05);下调miR-197的表达后,inhibitor组的CD82蛋白表达较其余两组显著增加(P<0.05);转染miR-197反义寡核苷酸的HCT116细胞的侵袭与迁移能力inhibitor组较NC组、Mock组均降低 (P<0.05)。结论 下调结肠癌HCT116细胞中miR-197的表达能抑制肿瘤细胞的侵袭和转移,其机制可能与其靶点CD82表达增加有关,这将为结肠癌的治疗提供新的靶点。

    Abstract:

    【Abstract】 Objective To investigate the effect of downregulating of microRNA-197 (miR-197) on the invasion and metastasis of colon cancer HCT116 cells and its mechanism. Methods HCT116 cells were randomly divided into miR-197 antisense oligonucleotide (inhibitor) group, negative control (NC) group and blank control (Mock) group. The miR-197 antisense oligonucleotide (miR-197 inhibitor) was transfected into HCT116 cells by liposome mediated transfection. The expression of miR-197 was determined by qRT-PCR. The target gene of miR-197 was predicted by bioinformatics, and the expression of CD82 protein was detected by Western blot. The invasion and migration ability of HCT116 cells was detected by transwell cell experiment. Results After transfection into HCT116 cells with miR-197 inhibitor, the expression of miR-197 in the inhibitor group was significantly lower than that of NC group and Mock group (P<0.05). After downregulating the expression of miR-197, the expression of CD82 protein in the inhibitor group was significantly higher than that of other two groups (P<0.05). The invasion and migration ability of HCT116 cells transfected with miR-197 inhibitor was lower than that of NC group and Mock group (P<0.05). Conclusion Downregulation of miR-197 in colon cancer HCT116 cells can inhibit invasion and metastasis of tumor cells, and the mechanism may be related to increased expression of the target CD82, which will provide a new target for the treatment of colon cancer.

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  • 在线发布日期: 2020-09-22
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