上海交通大学学报（医学版）

上海交通大学学报（医学版）

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miR-143在结肠癌组织中的差异表达及其对结肠癌细胞凋亡和迁移的影响

张晓云 1, 2，王立峰 1   

  1. 1. 上海交通大学 医学院附属新华医院，上海 200092；2. 上海中医药大学附属龙华医院病理科，上海 200032
  • 出版日期:2017-03-28 发布日期:2017-03-30
  • 通讯作者: 王立峰，电子信箱：wlf6009@163.com。
  • 作者简介:张晓云（1972—），女，副主任医师，硕士生；电子信箱：xiaoyunzhang1028@163.com。
  • 基金资助:

    上海中医药大学预算内基金项目（2013JW34）

Differential expression of miR-143 in colon cancer tissue and its effect on apoptosis and migration of colon cancer cell

ZHANG Xiao-yun1, 2, WANG Li-feng1   

  1. 1. Xinhua Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200092, China; 2. Department of Pathology, Longhua Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 200032, China
  • Online:2017-03-28 Published:2017-03-30
  • Supported by:

    Project of Shanghai University of Traditional Chinese Medicine, 2013JW34

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摘要：

目的 ·研究微小RNA-143（miR-143）在人结肠癌及癌周正常组织中的表达，探索其对结肠癌HCT116、RKO细胞功能的影响及下游靶基因的预测。方法 ·设计合成并构建miR-143慢病毒表达载体，稳定转染人结肠癌HCT116、RKO细胞。实时荧光定量PCR检测人结肠癌、癌周正常组织、慢病毒感染后HCT116和RKO细胞中miR-143的表达。Annexin V-APC单染法及流式细胞仪检测感染前后细胞凋亡能力。Transwell法检测感染前后HCT116、RKO细胞迁移能力的改变。Western blotting 方法检测FOSL2蛋白的表达。结果 · miR-143在结肠癌组织中的表达量显著低于癌周正常组织（0.339±0.454 vs 1.003±0.003）（U=16.000，Z=-4.231，P=0.000）。上调miR-143后，HCT116细胞凋亡率显著高于阴性对照组（P=0.000），RKO细胞凋亡率显著高于阴性对照组（P=0.000）。上调
miR-143后，HCT116迁移细胞数显著低于阴性对照组（P=0.000）。miR-143下调后，RKO迁移细胞数显著高于阴性对照组（P=0.003）。miR143下调后，FOSL2蛋白表达量升高。结论 · miR-143在结肠癌组织中表达量显著下降。上调miR-143可促进结肠癌细胞凋亡，并抑制HCT116细胞迁移；下调miR-143可促进RKO细胞迁移；抑制miR-143功能可上调FOSL2蛋白表达。

关键词: 结肠癌, 微小RNA-143, 凋亡, 迁移, FOSL2

Abstract:

Objective · To study the expression of microRNA-143 (miR-143) in human colon cancer and adjacent normal tissues, and to explore its effect on the function of colon cancer cells and downstream target genes. Methods · Lentiviral expression vector of miR-143 was designed and used to stably transfect colon cancer cell lines HCT116 and RKO. Real-time PCR was used to detect the expression of miR-143 in colon cancer, adjacent normal tissues, HCT116 cells and RKO cells infected with lentivirus. The apoptosis ratio of HCT116 cells and RKO cells was detected by Annexin V-APC single staining and flow cytometry. The migration ability of HCT116 cells and RKO cells was detected by Transwell method. The expression of FOSL2 protein was detected by Western blotting assay. Results · The expression of miR-143 in colon cancer tissue was significantly lower than that in adjacent mucosal tissue (0.339±0.454 vs 1.003±0.003) (U=16.000, Z=-4.231, P=0.000). By up-regulation of miR-143, apoptosis rates were significantly higher in HCT116 and RKO cells than those in the negative controls (P=0.000). The count of migrating HCT116 cells in up-regulated group was significantly lower than that in the negative control group (P=0.000). The count of migrating RKO cells in down-regulated group was significantly higher than that in the negative control (P=0.003). By down-regulation of miR-143, the expression of FOSL2 protein was increased. Conclusion · The expression of miR-143 was significantly decreased in colon cancer tissues. Up-regulation of miR-143 can promote the apoptosis rate in both cell lines, and inhibit the migration ability of HCT116 cells. Down-regulation of miR-143 could promote the migration ability of RKO cells. By inhibiting miR-143, the expression of FOSL2 protein in colonic cancer cells would be increased.

Key words: colon cancer, miR-143, apoptosis, migration, FOSL2