MicroRNA-210对卵巢癌细胞生长及放射敏感性的影响
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摘要
目的:探讨MicroRNA-210(miR-210)在卵巢癌细胞生长过程中的作用及其与放射治疗敏感性的关系,阐明miR-210对卵巢癌发展和治疗的影响及可能的分子机制。方法:体外培养人卵巢癌细胞株OVCAR3和SKOV3,应用细胞转染法分别将miR-210mimic和抗miR-210抑制剂转染至OVCAR3和SKOV3细胞中,并利用Real-time PCR法进行鉴定,得到miR-210过表达和低表达卵巢癌细胞模型。将2种细胞分别分为对照组、miR-210过表达组和miR-210低表达组,采用MTT法检测各组细胞增殖活性;采用不同剂量(30、50和100Gy)电离辐射照射对照组和miR-210过表达组细胞,采用MTT法检测各组细胞增殖活性;采用Western blotting法检测50Gy照射剂量组卵巢癌细胞中凋亡相关蛋白表达水平。所有实验细胞培养采用3复孔,并重复3次。结果:与对照组比较,miR-210过表达组细胞增殖活性升高,miR-210低表达组细胞增殖活性降低(P<0.05);在给予电离辐射照射后,与对照组比较,miR-210过表达组细胞增殖活性升高(P<0.05);且凋亡相关蛋白BAX在2株细胞中表达水平均明显下降,而BCL-2表达水平均明显升高。结论:miR-210可促进卵巢癌细胞的生长,同时通过抑制凋亡作用降低卵巢癌细胞对放射治疗的敏感性。
Objective:To investigate the role of MicroRNA-210(miR-210)in the growth process of ovarian cancer cells and its relationship with radiosensitivity,and to elucidate the effect of miR-210 on the development and treatment of ovarian cancer the possible molecular mechanism.Methods:The human ovarian cancer OVCAR3 and SKOV3 cells were cultured in vitro,miR-210 mimic and anti miR-210 inhibitors were transfected into the human ovarian cancer cell lines OVCAR3 and SKOV3,respectively,by cell transfection,and identified by Real-time PCR method;miR-210 overexpression and low expression of ovarian cancer cell models were obtained.The two kinds of cells were divided into control group,miR-210 overexpression model group and miR-210 low expression model group,and the cell proliferation activity was detected by MTT.The cells in control group and miR-210 overexpression model group were irradiated with different doses(30,50,and 100 Gy)of ionizing radiation,and the cell proliferation activity in each group was detected by MTT method.Western blotting method was used to detect the expression levels of apoptosis-related proteins in the ovarian cancer cells exposed to 50 Gy radiation dose.All the experimental cells were cultured with three double holes and repeated three times.Results:Compared with control group,the proliferation activity of the cells in miR-210 overexpression ovarian cancer cells group was enhanced,and the proliferation activity of the cells in miR-210 low expression cells group was reduced(P<0.05).After ionizing radiation,the proliferation activity of ovarian cancer cells in miRNA-210 overexpression group was enhanced compared with control group(P<0.05);the expression levels of apoptosis-related protein BAX in OVCAR3 and SKOV3 cells were significantly decreased,while the expression level of BCL-2 were significantly increased.Conclusion:miR-210 can promote the growth of ovarian cancer cells,and reduce the sensitivity of ovarian cancer cells to radiation therapy by inhibiting the apoptosis.
Objective:To investigate the role of MicroRNA-210(miR-210)in the growth process of ovarian cancer cells and its relationship with radiosensitivity,and to elucidate the effect of miR-210 on the development and treatment of ovarian cancer the possible molecular mechanism.Methods:The human ovarian cancer OVCAR3 and SKOV3 cells were cultured in vitro,miR-210 mimic and anti miR-210 inhibitors were transfected into the human ovarian cancer cell lines OVCAR3 and SKOV3,respectively,by cell transfection,and identified by Real-time PCR method;miR-210 overexpression and low expression of ovarian cancer cell models were obtained.The two kinds of cells were divided into control group,miR-210 overexpression model group and miR-210 low expression model group,and the cell proliferation activity was detected by MTT.The cells in control group and miR-210 overexpression model group were irradiated with different doses(30,50,and 100 Gy)of ionizing radiation,and the cell proliferation activity in each group was detected by MTT method.Western blotting method was used to detect the expression levels of apoptosis-related proteins in the ovarian cancer cells exposed to 50 Gy radiation dose.All the experimental cells were cultured with three double holes and repeated three times.Results:Compared with control group,the proliferation activity of the cells in miR-210 overexpression ovarian cancer cells group was enhanced,and the proliferation activity of the cells in miR-210 low expression cells group was reduced(P<0.05).After ionizing radiation,the proliferation activity of ovarian cancer cells in miRNA-210 overexpression group was enhanced compared with control group(P<0.05);the expression levels of apoptosis-related protein BAX in OVCAR3 and SKOV3 cells were significantly decreased,while the expression level of BCL-2 were significantly increased.Conclusion:miR-210 can promote the growth of ovarian cancer cells,and reduce the sensitivity of ovarian cancer cells to radiation therapy by inhibiting the apoptosis.
引文
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[19]张萍,王敏,接智慧,等.miR-210在化疗耐药与化疗敏感卵巢浆液性癌组织中的检测及意义[J].中国医科大学学报,2014,43(6):487-492.
[20]Rai B,Bansal A,Patel FD,et al.Radiotherapy for ovarian cancers-redefining the role[J].Asian Pac J Cancer Prev,2014,15(12):4759-4763.
[21]刘书中,李子全,李政垚,等.外周血miR-21检测在胰腺癌中的意义研究进展[J].中国实验诊断学,2016,20(6):1038-1042.
[22]付霞霏,何援利,王雪峰,等.化疗性卵巢功能早衰动物模型及患者血清miR-21水平的变化[J].解放军医学杂志,2016,41(11):915-918.
[2]Dong X,Men X,Zhang W,et al.Advances in tumor markers of ovarian cancer for early diagnosis[J].Indian J Cancer,2014,51(3):72-76.
[3]De Felice F,Marchetti C,Di Mino A,et al.Recurrent Ovarian Cancer:The Role of Radiation Therapy[J].Int J Gynecol Cancer,2017,27(4):690-695.
[4]Liao Q,Zhang HM,Li HH,et al.A preliminary study on the radiation-resistance mechanism in ovarian cancer[J].J Cancer Res Ther,2013,9(1):22-24.
[5]Reddy KB.MicroRNA(miRNA)in cancer[J].Cancer Cell Int,2015,15:38.
[6]霍红日,杨智勇,朱玲莉.miRNA-193a在肺腺癌患者体内的表达水平及意义[J].国际检验医学杂志,2017,38(13):1773-1775.
[7]Wu Q,Yang Z,Shi Y,et al.MiRNAs in human cancers:the diagnostic and therapeutic implications[J].Curr Pharm Des,2014,20(33):5336-5347.
[8]Li Y,Fang Y,Liu Y,et al.MicroRNAs in ovarian function and disorders[J].J Ovarian Res,2015,8:51.
[9]Zhang S,Lu Z,Unruh AK,et al.Clinically relevant microRNAs in ovarian cancer[J].Mol Cancer Res,2015,13(3):393-401.
[10]Liu M,Zhang X,Hu CF,et al.MicroRNA-mRNA functional pairs for cisplatin resistance in ovarian cancer cells[J].Chin J Cancer,2014,33(6):285-294.
[11]Su Z,Yang Z,Xu Y,et al.MicroRNAs in apoptosis,autophagy and necroptosis[J].Oncotarget,2015,6(11):8474-8490.
[12]Wang ZH,Xu CJ.Research Progress of MicroRNA in Early Detection of Ovarian Cancer[J].Chin Med J(Engl),2015,128(24):3363-3370.
[13]Gao YC,Wu J.MicroRNA-200c and microRNA-141 as potential diagnostic and prognostic biomarkers for ovarian cancer[J].Tumour Biol,2015,36(6):4843-4850.
[14]Shapira I,Oswald M,Lovecchio J,et al.Circulating biomarkers for detection of ovarian cancer and predicting cancer outcomes[J].Br J Cancer,2014,110(4):976-983.
[15]Mahdian-Shakib A,Dorostkar R,Tat M,et al.Differential role of microRNAs in prognosis,diagnosis,and therapy of ovarian cancer[J].Biomed Pharmacother,2016,84:592-600.
[16]Pal MK,Jaiswar SP,Dwivedi VN,et al.MicroRNA:a new and promising potential biomarker for diagnosis and prognosis of ovarian cancer[J].Cancer Biol Med,2015,12(4):328-341.
[17]Liu D,Xia H,Wang F,et al.MicroRNA-210interacts with FBXO31 to regulate cancer proliferation cell cycle and migration in human breast cancer[J].Onco Targets Ther,2016,9:5245-5255.
[18]Li L,Huang K,You Y,et al.Hypoxia-induced miR-210in epithelial ovarian cancer enhances cancer cell viability via promoting proliferation and inhibiting apoptosis[J].Int J Oncol,2014,44(6):2111-2120.
[19]张萍,王敏,接智慧,等.miR-210在化疗耐药与化疗敏感卵巢浆液性癌组织中的检测及意义[J].中国医科大学学报,2014,43(6):487-492.
[20]Rai B,Bansal A,Patel FD,et al.Radiotherapy for ovarian cancers-redefining the role[J].Asian Pac J Cancer Prev,2014,15(12):4759-4763.
[21]刘书中,李子全,李政垚,等.外周血miR-21检测在胰腺癌中的意义研究进展[J].中国实验诊断学,2016,20(6):1038-1042.
[22]付霞霏,何援利,王雪峰,等.化疗性卵巢功能早衰动物模型及患者血清miR-21水平的变化[J].解放军医学杂志,2016,41(11):915-918.