Li Rui, Shi Xuejun, Ling Fengyu, Wang Chunguang, Liu Junxia, Wang Wei, Li Ming
Department of Oncology, Yongchuan Hospital Affiliated to Chongqing Medical University, No. 439, Xuanhua Road, Chongqing, 402160, China.
Tumour Biol. 2015 Sep;36(9):7277-83. doi: 10.1007/s13277-015-3445-8. Epub 2015 Apr 21.
Increasing evidence has suggested that dysregulation of microRNAs (miRNAs) could contribute to tumor progression. The miR-34 family is directly transactivated by tumor suppressor p53 which is frequently mutated in various cancers; however, the effect of miR-34a on the ovarian cancer cells remains unclear. The aim of the paper was to study the expression of miR-34a in ovarian cancer and miR-34a's relation to the cell proliferation and metastasis in ovarian cancer in vitro. miR-34a expression was determined by quantitative RT-PCR in a panel of 60 human ovarian cancer samples. Functional characterization of miR-34a was accomplished by reconstitution of miR-34a expression in ovarian cancer cells by determining changes in proliferation, migration, and invasion. Our results showed that miR-34a is downregulated in ovarian cancer tissues compared with the corresponding adjacent non-neoplastic tissues, and the expression level of miR-34a was significantly lower in ovarian cancer cell lines in comparison with normal human fallopian tube epithelial cell line. The 3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-diphenytetrazoliumromide (MTT) assay revealed significant cell proliferation inhibition in miR-34a transfectant compared with the control from HO8910 and SKOV3 cells, which displayed lowest expressions of miR-34a. Furthermore, the transwell assay also showed significant cell migration inhibition in miR-34a transfectant, compared with cell lines transfected with NC. Overexpression of miR-34a led to the inhibition of AXL expression, indicating that AXL is a target gene for miR-34a. Our data suggest that miR-34a may function as a tumor suppressor through repression of oncogenic AXL in ovarian cancer.
越来越多的证据表明，微小RNA（miRNA）失调可能促进肿瘤进展。miR-34家族由肿瘤抑制因子p53直接反式激活，而p53在各种癌症中经常发生突变；然而，miR-34a对卵巢癌细胞的影响仍不清楚。本文旨在研究miR-34a在卵巢癌中的表达及其与卵巢癌细胞体外增殖和转移的关系。通过定量RT-PCR测定60例人卵巢癌样本中miR-34a的表达。通过测定miR-34a在卵巢癌细胞中表达的重建对细胞增殖、迁移和侵袭的影响，完成对miR-34a的功能表征。我们的结果显示，与相应的相邻非肿瘤组织相比，miR-34a在卵巢癌组织中表达下调，与正常人输卵管上皮细胞系相比，miR-34a在卵巢癌细胞系中的表达水平显著降低。3-（4,5）-二甲基噻唑（-z-y1）-3,5-二苯基四氮唑溴盐（MTT）试验显示，与来自HO8910和SKOV3细胞（miR-34a表达最低）的对照相比，miR-34a转染细胞中细胞增殖受到显著抑制。此外，与转染NC的细胞系相比，transwell试验也显示miR-34a转染细胞中细胞迁移受到显著抑制。miR-34a的过表达导致AXL表达受到抑制，表明AXL是miR-34a的靶基因。我们的数据表明，miR-34a可能通过抑制卵巢癌中致癌基因AXL的表达而发挥肿瘤抑制作用。
