Kawahara Kenta, Nagata Masashi, Yoshida Ryoji, Hirosue Akiyuki, Tanaka Takuya, Matsuoka Yuichiro, Arita Hidetaka, Nakashima Hikaru, Sakata Junki, Yamana Keisuke, Kawaguchi Sho, Gohara Shunsuke, Nagao Yuka, Hirayama Masatoshi, Takahashi Nozomu, Hirayama Mayumi, Nakayama Hideki
Department of Oral & Maxillofacial Surgery, Faculty of Life Sciences, Kumamoto University 1-1-1, Honjo, Chuo-ku, Kumamoto, 860-8556, Japan.
Department of Dentistry and Oral Surgery, Amakusa Central General Hospital, Amakusa 863-0033, Japan.
Biochem Biophys Rep. 2021 Sep 21;28:101114. doi: 10.1016/j.bbrep.2021.101114. eCollection 2021 Dec.
We aimed to determine the functional role of the miRNA, which affects drug sensitivity to 5-FU in oral squamous cell carcinoma (OSCC), using two types of 5-FU-resistant and parental OSCC cell lines. MiRNA microarray data showed that miR-30a was significantly upregulated in two resistant cell lines. Therefore, we investigated the effects and molecular mechanism of miR-30a on 5-FU sensitivity. Stable overexpression of miR-30a in parental OSCC cells decreased cell proliferation and attenuated drug sensitivity to 5-FU. Cell cycle analysis indicated that miR-30a overexpression increased the proportion of G1 phase cells and decreased the proportion of S phase cells. MiR-30a knockdown using siRNA reversed the effects of miR-30a overexpression. DNA microarray analysis using miR-30a-overexpressing cell lines and a TargetScan database search showed that is a target of miR-30a. A luciferase reporter assay confirmed that a miR-30a mimic interacted with the specific binding site in the 3' UTR of . knockdown with siRNA in OSCC cells yielded decreased drug sensitivity to 5-FU, similar to miR-30a overexpressing cells. These findings suggest that miR-30a in OSCC may be a novel biomarker of 5-FU-resistant tumors, as well as a therapeutic target for combating resistance.
我们旨在利用两种5-氟尿嘧啶（5-FU）耐药和亲本口腔鳞状细胞癌（OSCC）细胞系，确定影响口腔鳞状细胞癌对5-FU药物敏感性的微小RNA（miRNA）的功能作用。miRNA微阵列数据显示，miR-30a在两种耐药细胞系中显著上调。因此，我们研究了miR-30a对5-FU敏感性的影响及其分子机制。在亲本OSCC细胞中稳定过表达miR-30a可降低细胞增殖并减弱对5-FU的药物敏感性。细胞周期分析表明，miR-30a过表达增加了G1期细胞的比例，降低了S期细胞的比例。使用小干扰RNA（siRNA）敲低miR-30a可逆转miR-30a过表达的作用。使用过表达miR-30a的细胞系进行DNA微阵列分析以及搜索TargetScan数据库显示，[此处原文缺失相关基因名称]是miR-30a的一个靶标。荧光素酶报告基因检测证实，miR-30a模拟物与[此处原文缺失相关基因名称] 3'非翻译区（UTR）中的特异性结合位点相互作用。在OSCC细胞中用siRNA敲低[此处原文缺失相关基因名称]会导致对5-FU的药物敏感性降低，类似于过表达miR-30a的细胞。这些发现表明，OSCC中的miR-30a可能是5-FU耐药肿瘤的一种新型生物标志物，也是对抗耐药性的治疗靶点。
