Liu Bingyao, Cao Gang, Dong Zhen, Guo Ting
Department of Stomatology, Nanjing General Hospital, Nanjing, Jiangsu 210002, P.R. China.
Department of Endodontics, Nanjing Stomatological Hospital, Medical School of Nanjing University, Nanjing, Jiangsu 210008, P.R. China.
Oncol Lett. 2019 Jul;18(1):667-673. doi: 10.3892/ol.2019.10347. Epub 2019 May 13.
Effect of microRNA-27b (miR-27b) on cisplatin chemotherapy sensitivity of oral squamous cell carcinoma (OSCC) was investigated to provide a reference for clinical prevention of OSCC cell resistance. The clinical tissues of 34 patients with OSCC-resistant cancer and 28 patients with cisplatin-sensitive OSCC in Nanjing General Hospital were collected. The expression levels of miR-27b and Frizzled-7 (FZD7) in cancer tissues were detected by reverse transcription-quantitative polymerase chain reation (RT-qPCR). After a certain gradient of cisplatin was used to induce stable acquired resistance of OSCC cell line Tca8113/CDDP, a dose of miR-27b was added to construct a cell line overexpressing miR-27b in the drug-resistant cell line. The effect of cisplatin on the proliferation of drug-resistant OSCC cells was detected by colony formation assay. In addition, the scratch test and Transwell formation assay was performed to examine the effect of cisplatin drug stimulation on proliferation and migration of Tca8113/CDDP. Flow cytometry and Hoechst 33258 staining were used to detect the effect of miR-27b on apoptosis of OSCC-resistant cells after cisplatin chemotherapy. The expression level of miR-27b in cancer tissues of patients with drug-resistant OSCC was significantly lower than that of patients with OSCC cisplatin sensitivity (P<0.05). After high expression of miR-27b, the number of clones of drug-resistant OSCC cells after adding cisplatin drugs can be significantly inhibited. The proliferation and migration ability of drug-resistant OSCC was significantly decreased after the addition of cisplatin in miR-27b overexpression (P<0.05). miR-27 mimic enhanced the pro-apoptotic ability of cisplatin drugs (P<0.05). The expression of FZD7 in cisplation-resistant patients was significantly higher (P<0.05). miR-27b significantly inhibited the expression of FZD7 and β-catenin proteins. miR-27b can inhibit the resistance of OSCC to cisplatin drugs, increase apoptosis of cancer cells, and inhibit the proliferation of cancer cells. The mechanism may be related to the inhibition of FZD7/β-catenin signaling pathway activation in drug-resistant cell lines by miR-27b.
研究了微小RNA-27b（miR-27b）对口腔鳞状细胞癌（OSCC）顺铂化疗敏感性的影响，为临床预防OSCC细胞耐药提供参考。收集南京总医院34例OSCC耐药癌患者和28例顺铂敏感OSCC患者的临床组织。采用逆转录-定量聚合酶链反应（RT-qPCR）检测癌组织中miR-27b和卷曲蛋白7（FZD7）的表达水平。用一定梯度的顺铂诱导OSCC细胞系Tca8113/CDDP产生稳定的获得性耐药后，加入一定剂量的miR-27b，在耐药细胞系中构建过表达miR-27b的细胞系。采用集落形成试验检测顺铂对耐药OSCC细胞增殖的影响。此外，进行划痕试验和Transwell形成试验，以检测顺铂药物刺激对Tca8113/CDDP增殖和迁移的影响。采用流式细胞术和Hoechst 33258染色检测miR-27b对顺铂化疗后OSCC耐药细胞凋亡的影响。耐药OSCC患者癌组织中miR-27b的表达水平显著低于顺铂敏感OSCC患者（P<0.05）。miR-27b高表达后，加入顺铂药物后耐药OSCC细胞的克隆数可显著抑制。在miR-27b过表达的情况下，加入顺铂后，耐药OSCC的增殖和迁移能力显著降低（P<0.05）。miR-27模拟物增强了顺铂药物的促凋亡能力（P<0.05）。顺铂耐药患者中FZD7的表达显著更高（P<0.05）。miR-27b显著抑制FZD7和β-连环蛋白的表达。miR-27b可抑制OSCC对顺铂药物的耐药性，增加癌细胞凋亡，抑制癌细胞增殖。其机制可能与miR-27b抑制耐药细胞系中FZD7/β-连环蛋白信号通路的激活有关。
