Cao Ye, Zheng Jiajia, Lv Chentao
Department of General Surgery, Shanghai Public Health Clinical Center No. 921 Rd. Tongxin, Hongkou 200083 Shanghai China
Department of General Surgery, Zhongshan Hospital & Red Cross Hospital Xuhui 200030 Shanghai China.
RSC Adv. 2019 Jul 23;9(39):22755-22763. doi: 10.1039/c9ra01491h. eCollection 2019 Jul 17.
Dedifferentiated liposarcoma (DDLPS) is an aggressive tumor with high mortality. More insight into the biology of DDLPS tumorigenesis is needed to devise novel therapeutic approaches. Previous data showed that miRNA-199a-3p (miR-199a-3p) was strongly upregulated in DDLPS tissues. However, the biological role of miR-199a-3p in DDLPS remains unknown. In this study, we detected miR-199a-3p expression using RT-qPCR and observed that miR-199a-3p was more highly expressed in DDLPS tissues and cell lines (SW872 and LPS141). Functionally, MTT assay, flow cytometry and western blot results demonstrated that knockdown of miR-199a-3p inhibited DDLPS cell viability, enhanced apoptosis rate, and decreased expression of apoptosis-related genes Bax and cleaved caspase 3, as well as increased Bcl-2 expression . Moreover, xenograft tumors were generated and miR-199a-3p knockdown could suppress DDLPS xenograft tumor growth accompanying decreased proliferating cell nuclear antigen (PCNA) level and increased cleaved caspase 3 level . Mechanically, luciferase reporter assay and RNA immunoprecipitation (RIP) identified that CK1α was targeted and downregulated by miR-199a-3p. Expression of CK1α was lower in DDLPS tissues. Besides, there was a negative linear correlation between expressions of miR-199a-3p and CK1α in DDLPS tissues. Rescue experiments indicated that CK1α silencing could abolish the effect of miR-199a-3p knockdown on cell viability and apoptosis in DDLPS cells . In conclusion, knockdown of miR-199a-3p inhibits DDLPS cell viability and enhances apoptosis through targeting CK1α and . Our results suggest miR-199a-3p/CK1α axis may be a novel pathogen of DDLPS.
去分化脂肪肉瘤（DDLPS）是一种侵袭性肿瘤，死亡率很高。需要更深入了解DDLPS肿瘤发生的生物学机制，以设计新的治疗方法。先前的数据显示，miRNA-199a-3p（miR-199a-3p）在DDLPS组织中强烈上调。然而，miR-199a-3p在DDLPS中的生物学作用仍然未知。在本研究中，我们使用RT-qPCR检测miR-199a-3p表达，并观察到miR-199a-3p在DDLPS组织和细胞系（SW872和LPS141）中表达更高。在功能上，MTT法、流式细胞术和蛋白质印迹结果表明，敲低miR-199a-3p可抑制DDLPS细胞活力，提高凋亡率，降低凋亡相关基因Bax和裂解的caspase 3的表达，并增加Bcl-2的表达。此外，生成了异种移植肿瘤，敲低miR-199a-3p可抑制DDLPS异种移植肿瘤生长，同时增殖细胞核抗原（PCNA）水平降低，裂解的caspase 3水平升高。在机制上，荧光素酶报告基因检测和RNA免疫沉淀（RIP）确定CK1α是miR-199a-3p的靶标并被其下调。CK1α在DDLPS组织中的表达较低。此外，DDLPS组织中miR-199a-3p和CK1α的表达之间存在负线性相关。挽救实验表明，沉默CK1α可消除敲低miR-199a-3p对DDLPS细胞活力和凋亡的影响。总之，敲低miR-199a-3p通过靶向CK1α抑制DDLPS细胞活力并增强凋亡。我们的数据表明miR-199a-3p/CK1α轴可能是DDLPS的一种新发病机制。
