Yang Jianyu, Cui Ronghua, Liu Yingke
Workshop of National TCM Master, Sun Guangrong, The Harmonizing School of TCM, Beijing University of Chinese Medicine, Haikou, Hainan 570208, P.R. China.
Inheritance Workshop in Beijing Hepingli Hospital for National TCM Master, Sun Guangrong, Haikou, Hainan 570208, P.R. China.
Oncol Lett. 2020 Oct;20(4):23. doi: 10.3892/ol.2020.11884. Epub 2020 Jul 16.
Hepatocellular carcinoma (HCC) is one of the most common tumor malignances with poor chemotherapeutic efficiency due to chemoresistance. MicroRNAs (miRNAs) have essential roles in regulating chemoresistance. However, the mechanism underlying the involvement of miR-212-3p in paclitaxel (PTX) resistance in HCC remains unclear. PTX resistance was investigated in the present study by assessing cell viability, the half maximal inhibitory concentration of PTX, resistance-associated protein levels and apoptosis. The expression levels of miR-212-3p and zinc finger E-box binding homeobox 2 (ZEB2) were detected by reverse transcription-quantitative PCR and western blotting. The epithelial-mesenchymal transition (EMT), migration and invasion were evaluated by western blotting and transwell assay. The association between miR-212-3p and ZEB2 was investigating by the luciferase activity. The results showed that treatment of HCC cells with PTX inhibited cell viability and miR-212-3p level. Moreover, miR-212-3p was reduced and its overexpression resulted in decreased cell viability, half maximal inhibitory concentration (IC) of PTX and levels of P-glycoprotein and glutathione S-transferase π, but increased cell apoptosis, in Huh7/PTX cells. However, miR-212-3p knockdown induced opposite effects in Huh7 cells. Furthermore, EMT, migration and invasion were induced in Huh7/PTX cells and the addition of miR-212-3p inhibited EMT, migration and invasion. Meanwhile, miR-212-3p abrogation caused the opposite effects in Huh7 cells. Additionally, ZEB2 was directly targeted by miR-212-3p and its restoration or silencing abated the effect of miR-221-3p overexpression or knockdown in Huh7/PTX or Huh7 cells, respectively. The data from the present study suggest that miR-212-3p attenuates PTX resistance, by regulating EMT, migration and invasion via targeting ZEB2 in HCC cells, indicating a novel target for HCC chemotherapy.
肝细胞癌（HCC）是最常见的肿瘤恶性疾病之一，由于化疗耐药性，其化疗效率较低。微小RNA（miRNA）在调节化疗耐药性中起重要作用。然而，miR-212-3p参与HCC中紫杉醇（PTX）耐药性的潜在机制仍不清楚。本研究通过评估细胞活力、PTX的半数最大抑制浓度、耐药相关蛋白水平和细胞凋亡来研究PTX耐药性。通过逆转录定量PCR和蛋白质印迹法检测miR-212-3p和锌指E盒结合同源框2（ZEB2）的表达水平。通过蛋白质印迹法和Transwell实验评估上皮-间质转化（EMT）、迁移和侵袭。通过荧光素酶活性研究miR-212-3p与ZEB2之间的关联。结果表明，用PTX处理HCC细胞可抑制细胞活力和miR-212-3p水平。此外，在Huh7/PTX细胞中，miR-212-3p水平降低，其过表达导致细胞活力、PTX的半数最大抑制浓度（IC）以及P-糖蛋白和谷胱甘肽S-转移酶π水平降低，但细胞凋亡增加。然而，miR-212-3p敲低在Huh7细胞中诱导相反的效果。此外，Huh7/PTX细胞中诱导了EMT、迁移和侵袭，添加miR-212-3p可抑制EMT、迁移和侵袭。同时，miR-212-3p缺失在Huh7细胞中产生相反的效果。此外，ZEB2是miR-212-3p的直接靶点，其恢复或沉默分别减弱了miR-212-3p过表达或敲低对Huh7/PTX或Huh7细胞的影响。本研究数据表明，miR-212-3p通过靶向ZEB2调节HCC细胞中的EMT、迁移和侵袭，从而减弱PTX耐药性，为HCC化疗指明了一个新靶点。
