MicroRNA-98通过下调EZH2表达抑制结直肠癌细胞的活力与侵袭能力
|
摘要
目的:探讨微小RNA-98(mi R-98)与Zeste基因增强子同源物2(EZH2)之间的靶向关系,及其对结直肠癌细胞活力和侵袭能力的影响。方法:Target Scan软件预测EZH2和mi R-98之间的靶向结合,双萤光素酶报告基因实验验证mi R-98与EZH2之间的靶向关系。用mi R-98 mimic和mi R-98 inhibitor分别转染人结直肠癌SW480细胞和SW620细胞,Western blot检测EZH2的蛋白表达; MMT法检测细胞活力; Transwell法检测细胞的侵袭能力。转染EZH2过表达质粒检测其对细胞活力和侵袭的影响。结果:mi R-98能够靶向调节EZH2并负向调节SW480细胞和SW620细胞中EZH2的蛋白表达。在SW480细胞和SW620细胞中,过表达mi R-98显著下调细胞活力和侵袭能力,而抑制mi R-98表达显著促进细胞生长和侵袭。过表达EZH2也可促进SW480细胞和SW620细胞的生长和侵袭。结论:mi R-98通过下调EZH2表达抑制SW480细胞和SW620细胞的活力和侵袭。本研究为结直肠癌的治疗提供了新的靶点和方向。
AIM: To study the target relationship between micro RNA-98( mi R-98) and enhancer of Zeste homolog 2( EZH2),and the effect of mi R-98 on the viability and invasion ability of colorectal cancer cells. METHODS:The target relationship between EZH2 and mi R-98 was predicted by Target Scan software and confirmed by dual-luciferase reporter assay. The mi R-98 mimic and mi R-98 inhibitor were transfected into human colorectal cancer SW480 cells and SW620 cells. The protein expression level of EZH2 was determined by Western blot. The cell viability was measured by MTT assay,and the invasion ability was detected by Transwell assay. EZH2 over-expression vector was transfected into the colorectal cancer cells,and the cell viability and invasion ability were measured. RESULTS: mi R-98 targeted EZH2 and down-regulated EZH2 protein expression in the SW480 cells and SW620 cells. mi R-98 over-expression significantly decreased,while mi R-98 knockdown dramatically increased the viability and invasion ability of SW480 cells and SW620 cells. Additionally,EZH2 over-expression enhanced the viability and invasion ability of SW480 cells and SW620 cells.CONCLUSION: mi R-98 inhibits the viability and invasion ability of SW480 cells and SW620 cells by targeting EZH2,which may provide new therapeutic target and method for colorectal cancer treatment.
AIM: To study the target relationship between micro RNA-98( mi R-98) and enhancer of Zeste homolog 2( EZH2),and the effect of mi R-98 on the viability and invasion ability of colorectal cancer cells. METHODS:The target relationship between EZH2 and mi R-98 was predicted by Target Scan software and confirmed by dual-luciferase reporter assay. The mi R-98 mimic and mi R-98 inhibitor were transfected into human colorectal cancer SW480 cells and SW620 cells. The protein expression level of EZH2 was determined by Western blot. The cell viability was measured by MTT assay,and the invasion ability was detected by Transwell assay. EZH2 over-expression vector was transfected into the colorectal cancer cells,and the cell viability and invasion ability were measured. RESULTS: mi R-98 targeted EZH2 and down-regulated EZH2 protein expression in the SW480 cells and SW620 cells. mi R-98 over-expression significantly decreased,while mi R-98 knockdown dramatically increased the viability and invasion ability of SW480 cells and SW620 cells. Additionally,EZH2 over-expression enhanced the viability and invasion ability of SW480 cells and SW620 cells.CONCLUSION: mi R-98 inhibits the viability and invasion ability of SW480 cells and SW620 cells by targeting EZH2,which may provide new therapeutic target and method for colorectal cancer treatment.
引文
[1]Lee CHA,Kong JC,Ismail H,et al.Systematic review and meta-analysis of objective assessment of physical fitness in patients undergoing colorectal cancer surgery[J].Dis Colon Rectum,2018,61(3):400-409.
[2]Fang W.Chemotherapy in patient with colon cancer after renal transplantation:a case report with literature review[J].Medicine(Baltimore),2018,97(5):e9678.
[3]Li H,Chang C,Shang Y,et al.A missense variant in EZH2 is associated with colorectal cancer risk in a Chinese population[J].Oncotarget,2017,8(55):94738-94742.
[4]吴雪雷,蔡耀武,庄志忠,等.EZH2对胃癌细胞核因子κB靶基因的调节作用[J].中国病理生理杂志,2015,31(12):2169-2175.
[5]张丽静,刘博,赵增仁,等.上调mi R-187表达对人结肠癌细胞株增殖活性的影响[J].中国病理生理杂志,2013,29(11):1946-1951.
[6]O’Brien SJ,Carter JV,Burton JF,et al.The role of the mi R-200 family in epithelial-mesenchymal transition in colorectal cancer:a systematic review[J].Int J Cancer,2018,142(12):2501-2511.
[7]Wu F,Mo Q,Wan X,et al.NEAT1/hsa-mir-98-5p/MAPK6 axis is involved in non-small-cell lung cancer development[J].J Cell Biochem,2017 Nov 2.doi:10.1002/jcb.26442.[Epub ahead of print]
[8]Liu J,Chen W,Chen Z,et al.The effects of micro RNA-98 inhibits cell proliferation and invasion by targeting STAT3 in nasopharyngeal carcinoma[J].Biomed Pharmacother,2017,93:869-878.
[9]Zhu W,Huang Y,Pan Q,et al.MicroRNA-98 suppress Warburg effect by targeting HK2 in colon cancer cells[J].Dig Dis Sci,2017,62(3):660-668.
[10]Alldredge JK,Eskander RN.EZH2 inhibition in ARID1Amutated clear cell and endometrioid ovarian and endometrioid endometrial cancers[J].Gynecol Oncol Res Pract,2017,4:17.
[11]Christofides A,Karantanos T,Bardhan K,et al.Epigenetic regulation of cancer biology and anti-tumor immunity by EZH2[J].Oncotarget,2016,7(51):85624-85640.
[12]Vilorio-Marques L,Martin V,Diez-Tascon C,et al.The role of EZH2 in overall survival of colorectal cancer:a meta-analysis[J].Sci Rep,2017,7(1):13806.
[13]林琳,许庆文,刘春安.EZH2和桩蛋白在结直肠癌组织的表达及其与临床病理参数及预后的关系[J].中华实验外科杂志,2013,30(7):1520-1521.
[14]Li P,Zhang X,Wang H,et al.MALAT1 is associated with poor response to oxaliplatin-based chemotherapy in colorectal cancer patients and promotes chemoresistance through EZH2[J].Mol Cancer Ther,2017,16(4):739-751.
[15]Trino S,Lamorte D,Caivano A,et al.MicroRNAs as new biomarkers for diagnosis and prognosis,and as potential therapeutic targets in acute myeloid leukemia[J].Int JMol Sci,2018,19(2):E460.
[16]Bi S,Chai L,Yuan X,et al.MicroRNA-98 inhibits the cell proliferation of human hypertrophic scar fibroblasts via targeting Col1A1[J].Biol Res,2017,50:22.
[17]Cheng R,Dang R,Zhou Y,et al.MicroRNA-98 inhibits TGF-beta1-induced differentiation and collagen production of cardiac fibroblasts by targeting TGFBR1[J].Hum Cell,2017,30(3):192-200.
[18]Sun C,Liu H,Guo J,et al.MicroRNA-98 negatively regulates myocardial infarction-induced apoptosis by downregulating Fas and caspase-3[J].Sci Rep,2017,7(1):7460.
[19]Tan H,Zhu G,She L,et al.MiR-98 inhibits malignant progression via targeting MTDH in squamous cell carcinoma of the head and neck[J].Am J Cancer Res,2017,7(12):2554-2565.
[20]Huang Y,Hong X,Hu J,et al.Targeted regulation of mi R-98 on E2F1 increases chemosensitivity of leukemia cells K562/A02[J].Onco Targets Ther,2017,10:3233-3239.
[2]Fang W.Chemotherapy in patient with colon cancer after renal transplantation:a case report with literature review[J].Medicine(Baltimore),2018,97(5):e9678.
[3]Li H,Chang C,Shang Y,et al.A missense variant in EZH2 is associated with colorectal cancer risk in a Chinese population[J].Oncotarget,2017,8(55):94738-94742.
[4]吴雪雷,蔡耀武,庄志忠,等.EZH2对胃癌细胞核因子κB靶基因的调节作用[J].中国病理生理杂志,2015,31(12):2169-2175.
[5]张丽静,刘博,赵增仁,等.上调mi R-187表达对人结肠癌细胞株增殖活性的影响[J].中国病理生理杂志,2013,29(11):1946-1951.
[6]O’Brien SJ,Carter JV,Burton JF,et al.The role of the mi R-200 family in epithelial-mesenchymal transition in colorectal cancer:a systematic review[J].Int J Cancer,2018,142(12):2501-2511.
[7]Wu F,Mo Q,Wan X,et al.NEAT1/hsa-mir-98-5p/MAPK6 axis is involved in non-small-cell lung cancer development[J].J Cell Biochem,2017 Nov 2.doi:10.1002/jcb.26442.[Epub ahead of print]
[8]Liu J,Chen W,Chen Z,et al.The effects of micro RNA-98 inhibits cell proliferation and invasion by targeting STAT3 in nasopharyngeal carcinoma[J].Biomed Pharmacother,2017,93:869-878.
[9]Zhu W,Huang Y,Pan Q,et al.MicroRNA-98 suppress Warburg effect by targeting HK2 in colon cancer cells[J].Dig Dis Sci,2017,62(3):660-668.
[10]Alldredge JK,Eskander RN.EZH2 inhibition in ARID1Amutated clear cell and endometrioid ovarian and endometrioid endometrial cancers[J].Gynecol Oncol Res Pract,2017,4:17.
[11]Christofides A,Karantanos T,Bardhan K,et al.Epigenetic regulation of cancer biology and anti-tumor immunity by EZH2[J].Oncotarget,2016,7(51):85624-85640.
[12]Vilorio-Marques L,Martin V,Diez-Tascon C,et al.The role of EZH2 in overall survival of colorectal cancer:a meta-analysis[J].Sci Rep,2017,7(1):13806.
[13]林琳,许庆文,刘春安.EZH2和桩蛋白在结直肠癌组织的表达及其与临床病理参数及预后的关系[J].中华实验外科杂志,2013,30(7):1520-1521.
[14]Li P,Zhang X,Wang H,et al.MALAT1 is associated with poor response to oxaliplatin-based chemotherapy in colorectal cancer patients and promotes chemoresistance through EZH2[J].Mol Cancer Ther,2017,16(4):739-751.
[15]Trino S,Lamorte D,Caivano A,et al.MicroRNAs as new biomarkers for diagnosis and prognosis,and as potential therapeutic targets in acute myeloid leukemia[J].Int JMol Sci,2018,19(2):E460.
[16]Bi S,Chai L,Yuan X,et al.MicroRNA-98 inhibits the cell proliferation of human hypertrophic scar fibroblasts via targeting Col1A1[J].Biol Res,2017,50:22.
[17]Cheng R,Dang R,Zhou Y,et al.MicroRNA-98 inhibits TGF-beta1-induced differentiation and collagen production of cardiac fibroblasts by targeting TGFBR1[J].Hum Cell,2017,30(3):192-200.
[18]Sun C,Liu H,Guo J,et al.MicroRNA-98 negatively regulates myocardial infarction-induced apoptosis by downregulating Fas and caspase-3[J].Sci Rep,2017,7(1):7460.
[19]Tan H,Zhu G,She L,et al.MiR-98 inhibits malignant progression via targeting MTDH in squamous cell carcinoma of the head and neck[J].Am J Cancer Res,2017,7(12):2554-2565.
[20]Huang Y,Hong X,Hu J,et al.Targeted regulation of mi R-98 on E2F1 increases chemosensitivity of leukemia cells K562/A02[J].Onco Targets Ther,2017,10:3233-3239.