Lu Wuhao, Feng Long, Zhang Yan, Ma Yunyun, Li Ping, Wang Yuanyuan, Du Yuwen, Chen Xiaonan, Wu Shujun, Zhao Guoqiang, Lou Weihua
Department of Otolaryngology-Head and Neck Surgery, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan 450052, P.R. China.
Department of Pathogenic Organism Biology, Henan University of Chinese Medicine, Zhengzhou, Henan 450008, P.R. China.
Oncol Rep. 2016 Oct;36(4):2169-76. doi: 10.3892/or.2016.5049. Epub 2016 Aug 25.
Aberrant expression of miR-15a was recently reported in several types of cancers; however, its role in HPV-positive hypopharyngeal squamous cell carcinoma (HSCC) remains obscure. In the present study, we investigated the mechanism by which miR-15a induces HPV-positive HSCC apoptosis. Synthetic miR-15a mimics were transfected into FaDu cells (HPV-negative), and the miR-15a inhibitor was transfected into HPV-positive HSCC cells. miR-15a expression was analyzed by RT-PCR, and BCL2L2 and BCL2 were analyzed by western blotting. The Hochest 33342/propidium iodide (PI) and caspase-3/-9 assays, and Annexin V staining were used to assess the effect of miR-15a on apoptosis. After transfection, overexpression of miR-15a in the FaDu cells was associated with significantly decreased BCL2L2 and BCL2 expression and a significant increase in the apoptosis rate. The opposite results were observed in HPV-positive HSCC, where downregulation of miR-15a suppressed apoptosis. These findings indicate that miR-15a acts as a tumor suppressor in HPV-positive HSCC.
最近有报道称，miR-15a在多种癌症中存在异常表达；然而，其在人乳头瘤病毒（HPV）阳性的下咽鳞状细胞癌（HSCC）中的作用仍不清楚。在本研究中，我们探究了miR-15a诱导HPV阳性HSCC细胞凋亡的机制。将合成的miR-15a模拟物转染至FaDu细胞（HPV阴性），并将miR-15a抑制剂转染至HPV阳性的HSCC细胞。通过逆转录聚合酶链反应（RT-PCR）分析miR-15a的表达，并通过蛋白质免疫印迹法分析BCL2L2和BCL2。采用Hochest 33342/碘化丙啶（PI）、半胱天冬酶-3/-9检测以及膜联蛋白V染色来评估miR-15a对细胞凋亡的影响。转染后，FaDu细胞中miR-15a的过表达与BCL2L2和BCL2表达显著降低以及凋亡率显著升高相关。在HPV阳性的HSCC中观察到相反的结果，其中miR-15a的下调抑制了细胞凋亡。这些发现表明，miR-15a在HPV阳性的HSCC中起肿瘤抑制作用。
