miRNA-143 靶向MACC1 抑制宫颈癌细胞侵袭

目的：探讨miRNA-143 对宫颈癌细胞侵袭能力的影响。方法：采用脂质体转染法瞬时转染miRNA-143 过表达和干扰质粒，Transwell 迁移实验检测miRNA-143 过表达和抑制后宫颈癌细胞侵袭能力的改变，生物信息学预测miRNA-143 的作用靶点。miRNA-143 过表达和抑制后Westernblot及双荧光素酶报告基因检测其靶点MACC1 表达，RT-qPCR检测20例患者宫颈癌和癌旁正常组织标本中miRNA-143 和MACC1 mRNA 的表达，分析20例患者宫颈癌组织中miRNA-143 和MACC1 mRNA 表达的相关性。结果：Transwell 迁移实验显示miRNA-143 过表达的宫颈癌细胞的侵袭能力降低，抑制miRNA-143 后侵袭能力增强。生物信息学预测显示miRNA-143 作用于MACC1 的3'-UTR ，Westernblot及双荧光素酶报告基因结果进一步证实miRNA-143 作用于MACC1 的3'-UTR 。RT-qPCR显示miRNA-143 过表达的MACC1 mRNA 表达下降，而抑制miRNA-143 后MACC1 mRNA 表达上升。抑制miRNA-143 表达的宫颈癌细胞中MACC1 被干扰后，宫颈癌细胞的侵袭能力显著被抑制。宫颈癌组织中miRNA-143表达水平显著低于正常宫颈上皮组织，MACC1 表达水平显著高于正常宫颈上皮组织，20例患者的宫颈癌组织中miRNA-143 与MACC1 mRNA 表达呈负相关。结论：miRNA-143 在宫颈癌中表达水平下降，并可能通过靶向MACC1 调节宫颈癌细胞的侵袭能力。

Inhibitory effect of miRNA-143 on the invasiveness of cervical cancer cells by targeting MACC1

Objective:To illustrate the role of miRNA-143 on the invasiveness of cervical cancer cells.Methods:MiRNA-143 mimics or inhibitor sequences were transiently expressed in the cervical cancer cells by liposome transfection. Transwell assay was ap-plied to test the invasive ability of cervical cancer cells after miRNA-143 over-expression or inhibition. Bioinformatics assay was used to predict the targets of miRNA- 143. RT- qPCR and luciferase reporter assay were performed to detect the expression of MACC 1 mRNA in the cancer cells. RT-qPCR was conducted to test the expression of miRNA-143 and MACC 1 mRNA in 20fresh primary cervi -cal cancer and their matched para-neoplastic tissues. Statistical analyses were performed to evaluate the association between the expres -sion of miRNA-143 and MACC 1 mRNA in the 20cases of cervical cancer. Results: Transwell assays revealed that the miRNA- 143 over-expression inhibited the cell invasiveness, while miRNA- 143 inhibition promoted the invasive ability of the cervical cancer cells. Bioinformatics analyses revealed that miRNA-143 could target the 3'- UTR of MACC1. Dual luciferase reporter assay confirmed that miRNA-143 can affect 3'-UTR sequence in MACC 1 genes. RT-qPCR analyses indicated that the expression of MACC1 mRNA was ob -viously down-regulated after miRNA-143 over-expression, while significantly increased after the miRNA- 143 inhibition. The migration in Caski/miRNA-143 inhibitor cells was obviously elevated after being transfected with MACC1 shRNAs. RT-qPCR analyses showed that the expression of miRNA- 143 was obviously decreased in the cancer tissues compared with the normal tissues, while MACC1 mRNA was apparently decreased in cancer tissues compared with the normal ones. Statistical analyses revealed that miRNA-143 was negatively correlated with MACC 1 mRNA in the20cases of cervical cancer. Conclusion:This study reveals that miRNA- 143 is down-regulated in the cervical cancer tissues. MiRNA- 143 may play an important role in the regulation of cell invasiveness by targeting MACC 1 in the cervical cancer cells.