miR-137对肾癌GRC-1细胞增殖与凋亡的影响
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摘要
目的研究miR-137对肾癌GRC-1细胞增殖与凋亡的影响。方法采用miR-137模拟物转染至肾癌GRC-1细胞,实验分为未转染组(未进行miR-137模拟物转染)、miR-137对照组(转染随机序列)、miR-137转染组(进行miR-137模拟物转染)。荧光定量PCR检测各组转染后48h细胞中miR-137表达水平,应用噻唑蓝细胞增殖试验(MTT)、流式细胞检测技术与免疫荧光评价miR-137对肾癌GRC-1细胞增殖和凋亡的影响。结果转染miR-137模拟物48h后,荧光定量PCR检测发现未转染组、miR-137对照组及miR-137转染组GRC-1细胞中miR-137的相对表达量依次为(24.43±2.03)%、(26.57±2.55)%、和(73.30±3.29)%,差异有统计学意义(P<0.05)。MTT细胞增殖实验与流式细胞仪检测结果显示,与未转染组、miR-137对照组相比,miR-137转染组转染48h后肾癌GRC-1细胞的增殖率显著降低,凋亡率显著升高,差异有统计学意义(P<0.05),而未转染组、miR-137对照组肾癌GRC-1细胞的增殖率、凋亡率比较差异无统计学意义(P>0.02)。结论 miR-137可明显抑制肾癌GRC-1细胞增殖和促进其凋亡,故有望成为肾癌基因治疗的新靶点。
Objective To study the effect of miR-137 on proliferation and apoptosis of renal carcinoma GRC-1 cells. Methods miR-137 analogies were transfected into renal carcinoma GRC-1 cells. The non-transfection group( without transfection of miR-137 analogies),the miR-137 control group( transfected with random sequence) and the miR-137 transfection group( with transfection of miR-137 analogies) were used. The expression of miR-137 in cells was detected by fluorescence quantitative PCR in 48 h after transfection. The effect of miR-137 on proliferation and apoptosis of renal carcinoma GRC-1 cells was evaluated with the methyl-thiazolyl-tetrazolium( MTT) test,flowcytometry and immunofluorescence. Results 48 hours after the transfection of miR-137 analogies,fluorescence quantitative PCR detected that the relative expression levels of miR-137 in GRC-1 cells in the non-transfection group,the miR-137 control group and the miR-137 transfection group were( 24. 43 ± 2. 03) %,( 26. 57 ± 2. 55) % and( 73. 30 ± 3. 29) %,respectively( P <0. 05). MTT cell proliferation assay and flowcytometry showed that the proliferation rate of renal carcinoma GRC-1 cells was significantly lower and the apoptosis rate was significantly higher in the miR-137 transfection group than the non-transfection group and miR-137 control group at 48 hours after transfection( P < 0. 05). However,there was no significant difference between the non-transfection group and miR-137 control group in the proliferation rate and apoptosis rate of renal carcinoma GRC-1 cells( P > 0. 02).Conclusion miR-137 can inhibit the proliferation and promote the apoptosis of GRC-1 cells,which might be a newtarget for gene therapy of renal carcinoma.
Objective To study the effect of miR-137 on proliferation and apoptosis of renal carcinoma GRC-1 cells. Methods miR-137 analogies were transfected into renal carcinoma GRC-1 cells. The non-transfection group( without transfection of miR-137 analogies),the miR-137 control group( transfected with random sequence) and the miR-137 transfection group( with transfection of miR-137 analogies) were used. The expression of miR-137 in cells was detected by fluorescence quantitative PCR in 48 h after transfection. The effect of miR-137 on proliferation and apoptosis of renal carcinoma GRC-1 cells was evaluated with the methyl-thiazolyl-tetrazolium( MTT) test,flowcytometry and immunofluorescence. Results 48 hours after the transfection of miR-137 analogies,fluorescence quantitative PCR detected that the relative expression levels of miR-137 in GRC-1 cells in the non-transfection group,the miR-137 control group and the miR-137 transfection group were( 24. 43 ± 2. 03) %,( 26. 57 ± 2. 55) % and( 73. 30 ± 3. 29) %,respectively( P <0. 05). MTT cell proliferation assay and flowcytometry showed that the proliferation rate of renal carcinoma GRC-1 cells was significantly lower and the apoptosis rate was significantly higher in the miR-137 transfection group than the non-transfection group and miR-137 control group at 48 hours after transfection( P < 0. 05). However,there was no significant difference between the non-transfection group and miR-137 control group in the proliferation rate and apoptosis rate of renal carcinoma GRC-1 cells( P > 0. 02).Conclusion miR-137 can inhibit the proliferation and promote the apoptosis of GRC-1 cells,which might be a newtarget for gene therapy of renal carcinoma.
引文
[1]刘会锋,闫泽晨,顾朝辉,等.微小RNA-143抑制人肾癌细胞株A498增殖及其对凋亡的影响.中华实验外科杂志,2013,30(3):577-578.
[2]罗锦花,田亚平.从血常规分析癌症患者血细胞参数的变化.标记免疫分析与临床,2013,20(3):188-190.
[3]王国栋,党亚正.PIRH2在肾细胞癌中的表达及其意义.西部医学,2014,26(3):280-282.
[4]李栋,刘希胜,王新明,等.miR-143对肾癌GRC-1细胞增殖与凋亡及其靶点HIF-1α的影响.临床肿瘤学杂志,2016,21(11):967-971.
[5]Sang L,Liu T,Liu H,et al.MicroRNA-137 suppresses cell migration and invasion in renal cell carcinoma by targeting PIK3R3.Int J Clin Exp Med,2016,9(4):7160-7167.
[6]陈美源,肖杰,江建新,等.MicroRNA-137抑制胰腺癌细胞侵袭和迁移能力.贵阳医学院学报,2015,40(2):112-116,120.
[7]俞能旺,张爱民,郝俊文,等.miR-21对肾癌细胞凋亡和侵袭力的影响.现代生物医学进展,2013,13(4):636-638.
[8]常永梅,王明智,孙聪,等.AEG-1抑制miR-137对非小细胞肺癌增殖调控作用.中南医学科学杂志,2016,44(3):279-282.
[9]林丽娜,陈为,王伟民,等.miR-137对U87胶质瘤细胞中EZH2表达和细胞增殖的影响.中国微侵袭神经外科杂志,2011,16(8):362-365.
[10]曹影,王培蓓,马荧雪,等.miR-137对人喉癌细胞株Hep-2细胞增殖的影响.江苏医药,2013,39(9):1015-1017.
[11]Neault M,Mallette F A,Richard S.miR-137modulates a tumor suppressor network-inducing senescence in pancreatic cancer cells.Cell Reports,2016,14(8):1966-1978.
[12]刘晓阳,郑海伦,任志,等.microRNA-137对人胃癌MKN-45细胞迁移侵袭的影响.安徽医科大学学报,2015,50(12):1748-1752.
[13]Zhao Y,Li Y,Lou G,et al.MiR-137 targets estrogen-related receptor alpha and impairs the proliferative and migratory capacity of breast cancer cells.PLo S One,2012,7(6):e39102.
[14]徐丽伟,漆靖,卢太亮,等.miR-137与胃癌侵袭能力的关系研究.中国普通外科杂志,2015,24(4):600-603.
[15]郭俊,李民,万政强,等.MiR-137对人脑胶质瘤细胞株U87细胞增殖和凋亡影响的体外研究.中华神经医学杂志,2013,12(12):1216-1219.
[16]Zhang H,Li H.miR-137 inhibits renal cell carcinoma growth in vitro and in vivo.Oncol Lett,2016,12(1):715-720.
[2]罗锦花,田亚平.从血常规分析癌症患者血细胞参数的变化.标记免疫分析与临床,2013,20(3):188-190.
[3]王国栋,党亚正.PIRH2在肾细胞癌中的表达及其意义.西部医学,2014,26(3):280-282.
[4]李栋,刘希胜,王新明,等.miR-143对肾癌GRC-1细胞增殖与凋亡及其靶点HIF-1α的影响.临床肿瘤学杂志,2016,21(11):967-971.
[5]Sang L,Liu T,Liu H,et al.MicroRNA-137 suppresses cell migration and invasion in renal cell carcinoma by targeting PIK3R3.Int J Clin Exp Med,2016,9(4):7160-7167.
[6]陈美源,肖杰,江建新,等.MicroRNA-137抑制胰腺癌细胞侵袭和迁移能力.贵阳医学院学报,2015,40(2):112-116,120.
[7]俞能旺,张爱民,郝俊文,等.miR-21对肾癌细胞凋亡和侵袭力的影响.现代生物医学进展,2013,13(4):636-638.
[8]常永梅,王明智,孙聪,等.AEG-1抑制miR-137对非小细胞肺癌增殖调控作用.中南医学科学杂志,2016,44(3):279-282.
[9]林丽娜,陈为,王伟民,等.miR-137对U87胶质瘤细胞中EZH2表达和细胞增殖的影响.中国微侵袭神经外科杂志,2011,16(8):362-365.
[10]曹影,王培蓓,马荧雪,等.miR-137对人喉癌细胞株Hep-2细胞增殖的影响.江苏医药,2013,39(9):1015-1017.
[11]Neault M,Mallette F A,Richard S.miR-137modulates a tumor suppressor network-inducing senescence in pancreatic cancer cells.Cell Reports,2016,14(8):1966-1978.
[12]刘晓阳,郑海伦,任志,等.microRNA-137对人胃癌MKN-45细胞迁移侵袭的影响.安徽医科大学学报,2015,50(12):1748-1752.
[13]Zhao Y,Li Y,Lou G,et al.MiR-137 targets estrogen-related receptor alpha and impairs the proliferative and migratory capacity of breast cancer cells.PLo S One,2012,7(6):e39102.
[14]徐丽伟,漆靖,卢太亮,等.miR-137与胃癌侵袭能力的关系研究.中国普通外科杂志,2015,24(4):600-603.
[15]郭俊,李民,万政强,等.MiR-137对人脑胶质瘤细胞株U87细胞增殖和凋亡影响的体外研究.中华神经医学杂志,2013,12(12):1216-1219.
[16]Zhang H,Li H.miR-137 inhibits renal cell carcinoma growth in vitro and in vivo.Oncol Lett,2016,12(1):715-720.