摘要
目的探讨瑞芬太尼对骨肉瘤细胞增殖、凋亡的影响及其作用机制。方法2017年8月—2018年12月于山东省枣庄矿业集团枣庄医院进行实验。收集经病理确诊的骨肉瘤患者36例,取其骨肉瘤组织及癌旁组织,采用实时荧光定量聚合酶链反应(qRT-PCR)检测其miR-148a的表达。以骨肉瘤MG63细胞为研究对象,设立对照组及瑞芬太尼0.1、1、10、100、200μmol/L组,分别处理MG63细胞,采用MTT细胞增殖实验测定瑞芬太尼抑制MG63细胞增殖的效果;流式细胞术检测瑞芬太尼对MG63细胞凋亡率的影响;qRT-PCR检测瑞芬太尼对MG63细胞中miR-148a表达的影响。将MG63细胞随机分为瑞芬太尼10μmol/L+anti-miR-NC亚组与瑞芬太尼10μmol/L+anti-miR-148a亚组,观测抑制miR-148a表达验证瑞芬太尼对MG63细胞增殖及凋亡的作用机制。Western-blot检测Cyclin D1、P21 Bax、Bcl-2蛋白表达。结果与癌旁组织比较,骨肉瘤组织中miR-148a的表达水平显著降低(t/P=20.402/<0.001);24 h、48 h、72 h时瑞芬太尼各组可抑制骨肉瘤细胞增殖(F=182.419,189.181,165.061,P均<0.001),促进细胞凋亡(F/P=110.203/<0.001),随瑞芬太尼浓度升高P21表达水平升高(F/P=355.973/<0.001),Cyclin D1表达水平降低(F/P=273.731/<0.001),并可明显促进Bax表达(F/P=156.941/<0.001),抑制Bcl-2表达(F/P=288.568/<0.001);与瑞芬太尼10μmol/L+anti-miR-NC亚组比较,瑞芬太尼10μmol/L+anti-miR-148a亚组MG63细胞增殖活力显著升高,细胞凋亡率显著升高(P<0.01)。结论瑞芬太尼可能通过上调miR-148a表达诱导骨肉瘤细胞凋亡,同时具有抑制骨肉瘤细胞增殖的作用。
Objective To investigate the effect of remifentanil on proliferation and apoptosis of osteosarcoma cells and its mechanism.Methods The experiment was conducted in Zaozhuang Hospital of Shandong Zaozhuang Mining Group from August 2017 to December 2018.The expression of Mir-148A was detected by real-time fluorescent quantitative polymerase chain reaction(qRT-PCR).The control group and remifentanil 0.1,1,10,100,200μmol/L group were established.MG63 cells were treated with remifentanil respectively,and the inhibitory effect of remifentanil on MG63 cell proliferation was measured by MTT cell proliferation experiment.The effect of remifentanil on the apoptosis rate of MG63 cells was detected by flow cytometry,and the expression of miR-148a in MG63 cells was detected by qRT-PCR.MG63 cells were randomly divided into remifentanil 10μmol/L+anti-miR-NC sub group and remifentanil 10μmol/L+anti-miR-148a sub group.The inhibition of miR-148a expression was observed to verify the mechanism of remifentanil on the proliferation and apoptosis of MG63 cells.Western blot was used to detect the expression of cyclin D1,P21 Bax and Bcl-2.Results The expression level of miR-148a in osteosarcoma was significantly lower than that in paracancerous tissues(t/P=20.402/<0.001).At 24 h,48 h and 72 h,remifentanil could inhibit the proliferation of osteosarcoma cells(F=182.419,189.181,165.061,P<0.001),promote apoptosis(F/P=110.203/<0.001),and increase p21 expression level(F/P=355.973/<0.001),cyclin The expression level of D1 decreased(F/P=273.731/<0.001),and it could promote the expression of Bax(F/P=156.941/<0.001),and inhibit the expression of Bcl 2(F/P=288.568/<0.001).Compared with remifentanil 10μmol/L+anti-miR-NC subgroup,the proliferation activity and apoptosis rate of MG63 cells in remifentanil 10μmol/L+anti-miR-148A subgroup increased significantly(P<0.01).Conclusion Remifentanil may induce apoptosis of osteosarcoma cells by up regulating the expression of miR-148a and inhibit the proliferation of osteosarcoma cells.
作者
孟杨
王绍伟
王宪峰
徐海伟
Meng Yang;Wang Shaowei;Wang Xianfeng;Xu Haiwei(Department of Orthopaedics, Zaozhuang Hospital, Zaozhuang Mining Group, Shandong Province,Zaozhuang 277100,China)
出处
《疑难病杂志》
CAS
2020年第4期371-375,共5页
Chinese Journal of Difficult and Complicated Cases
作者简介
通信作者:王宪峰,E-mail:wtswn19@163.com。
