上调miRNA-27a-3p对乳腺癌MCF-7细胞增殖、侵袭和迁移能力的影响
|
摘要
目的:探讨上调miRNA-27a-3p(miR-27a-3p)对人乳腺癌MCF-7细胞增殖、侵袭和迁移能力的影响。方法:将miR-27a-3p模拟物转染至MCF-7细胞中,以转染模拟物对照的细胞为阴性对照,以不转染的细胞为空白对照。通过qRT-PCR检测miR-27a-3p的表达,CCK-8实验检测细胞增殖情况,Transwell实验检测细胞侵袭和迁移能力,Western blot检测细胞中PCNA、MMP-2、MMP-9蛋白的表达。结果:与空白对照组和阴性对照组相比,模拟物组MCF-7细胞中miR-27a-3p表达水平,细胞增殖、侵袭和迁移能力,PCNA、MMP-2和MMP-9蛋白表达水平均升高(P <0. 05)。结论:上调miR-27a-3p能够提高MCF-7细胞的增殖、侵袭和迁移能力,其机制可能与促进PCNA、MMP-2和MMP-9蛋白表达有关。
Aim: To investigate the effects of up-regulation of miRNA-27 a-3 p( miR-27 a-3 p) on proliferation,invasion and migration of human breast cancer MCF-7 cells. Methods: MCF-7 cells were cultured in vitro,and miR-27 a-3 p mimics were transfected into MCF-7 cells. The cells transfected mimics control were used as negative control,and the non-transfected cells were used as blank control. qRT-PCR was used to detect transfection efficiency. CCK-8 assay was used to detect the proliferation of MCF-7 cells. Transwell assay was used to detect the invasion and migration of MCF-7 cells. Western blot was used to detect the expressions of PCNA,MMP-2,and MMP-9 protein in MCF-7 cells. Results: Compared with the negative control group and the blank control group,miR-27 a-3 p expression level was significantly increased in miR-27 a-3 p mimics group,cell proliferation,invasion and migration were significantly prompted,and PCNA,MMP-2 and MMP-9 protein levels were significantly increased( P < 0. 05). Conclusion: Up-regulation of miR-27 a-3 p could promote the proliferation,invasion and migration of MCF-7 cells,which may be related with its promoting the expressions of PCNA,MMP-2 and MMP-9 proteins.
Aim: To investigate the effects of up-regulation of miRNA-27 a-3 p( miR-27 a-3 p) on proliferation,invasion and migration of human breast cancer MCF-7 cells. Methods: MCF-7 cells were cultured in vitro,and miR-27 a-3 p mimics were transfected into MCF-7 cells. The cells transfected mimics control were used as negative control,and the non-transfected cells were used as blank control. qRT-PCR was used to detect transfection efficiency. CCK-8 assay was used to detect the proliferation of MCF-7 cells. Transwell assay was used to detect the invasion and migration of MCF-7 cells. Western blot was used to detect the expressions of PCNA,MMP-2,and MMP-9 protein in MCF-7 cells. Results: Compared with the negative control group and the blank control group,miR-27 a-3 p expression level was significantly increased in miR-27 a-3 p mimics group,cell proliferation,invasion and migration were significantly prompted,and PCNA,MMP-2 and MMP-9 protein levels were significantly increased( P < 0. 05). Conclusion: Up-regulation of miR-27 a-3 p could promote the proliferation,invasion and migration of MCF-7 cells,which may be related with its promoting the expressions of PCNA,MMP-2 and MMP-9 proteins.
引文
[1]左婷婷,陈万青.中国乳腺癌全人群生存率分析研究进展[J].中国肿瘤临床,2016,43(14):639
[2]ATWAL D,RAMOS JM,MAKHOUL I.Late breast cancer recurrence with bone marrow metastases and acute pulmonary hypertension[J].Proc(Bayl Univ Med Cent),2018,31(2):213
[3]ZHANG H,ZHANG Y,ZHAO X,et al.Association of two microRNA polymorphisms miR-27 rs895819 and miR-423rs6505162 with the risk of cancer[J].Oncotarget,2017,8(29):46969
[4]TANAKA K,MIYATA H,SUGIMURA K,et al.miR-27 is associated with chemoresistance in esophageal cancer through transformation of normal fibroblasts to cancer-associated fibroblasts[J].Carcinogenesis,2015,36(8):894
[5]魏疆,禄邵英.miR-27在胃癌中的表达及其作用研究[J].陕西医学杂志,2015,44(5):620
[6]SUN Y,YANG X,LIU M,et al.B4GALT3 up-regulation by miR-27a contributes to the oncogenic activity in human cervical cancer cells[J].Cancer Lett,2016,375(2):284
[7]LIU J,MAO Y,ZHANG D,et al.MiR-143 inhibits tumor cell proliferation and invasion by targeting STAT3 in esophageal squamous cell carcinoma[J].Cancer Lett,2016,373(1):97
[8]TANG W,ZHU J,SU S,et al.MiR-27 as a prognostic marker for breast cancer progression and patient survival[J].PLo S One,2012,7(12):e51702
[9]MCALLISTER SD,MURASE R,CHRISTIAN RT,et al.Pathways mediating the effects of cannabidiol on the reduction of breast cancer cell proliferation,invasion,and metastasis[J].Breast Cancer Res Treat,2011,129(1):37
[10]杨浚沨,王龙强,李海,等.乳腺癌循环miRNA生物标志物的筛选及验证[J].中国普通外科杂志,2015,24(5):696
[11]QIN W,ZHANG K,CLARKE K,et al.Methylation and miRNA effects of resveratrol on mammary tumors vs.normal tissue[J].Nutr Cancer,2014,66(2):270
[12]CELI-TERRASSA T,LIU DD,CHOUDHURY A,et al.Normal and cancerous mammary stem cells evade interferon-induced constraint through the miR-199a-LCOR axis[J].Nat Cell Biol,2017,19(6):711
[13]PENG H,WANG X,ZHANG P,et al.miR-27a promotes cell proliferation and metastasis in renal cell carcinoma[J].Int J Clin Exp Pathol,2015,8(2):2259
[14]ZHOU L,LIANG X,ZHANG L,et al.MiR-27a-3p functions as an oncogene in gastric cancer by targeting BTG2[J].Oncotarget,2016,7(32):51943
[15]WANG YL,GONG WG,YUAN QL.Effects of miR-27a upregulation on thyroid cancer cells migration,invasion,and angiogenesis[J].Genet Mol Res,2016,15(4):38
[16]WANG T,XU Z.miR-27 promotes osteoblast differentiation by modulating Wnt signaling[J].Biochem Biophys Res Commun,2010,402(2):186
[17]顾文元.利拉鲁肽对人乳腺癌MCF-7细胞增殖、凋亡的影响及其机制研究[D].天津:天津医科大学,2016.
[18]ZHOU J,LI K,GU Y,et al.Transcriptional up-regulation of Rho E by hypoxia-inducible factor(HIF)-1 promotes epithelial to mesenchymal transition of gastric cancer cells during hypoxia[J].Biochem Biophys Res Commun,2011,415(2):348
[19]CHOI BD,JEONG SJ,WANG G,et al.Secretory leukocyte protease inhibitor is associated with MMP-2 and MMP-9 to promote migration and invasion in SNU638 gastric cancer cells[J].Int J Mol Med,2011,28(4):527
[20]BURLAKA AP,GANUSEVICH II,GAFUROV MR,et al.Stomach cancer:interconnection between the redox state,activity of MMP-2,MMP-9 and stage of tumor growth[J].Cancer Microenviron,2016,9(1):27
[2]ATWAL D,RAMOS JM,MAKHOUL I.Late breast cancer recurrence with bone marrow metastases and acute pulmonary hypertension[J].Proc(Bayl Univ Med Cent),2018,31(2):213
[3]ZHANG H,ZHANG Y,ZHAO X,et al.Association of two microRNA polymorphisms miR-27 rs895819 and miR-423rs6505162 with the risk of cancer[J].Oncotarget,2017,8(29):46969
[4]TANAKA K,MIYATA H,SUGIMURA K,et al.miR-27 is associated with chemoresistance in esophageal cancer through transformation of normal fibroblasts to cancer-associated fibroblasts[J].Carcinogenesis,2015,36(8):894
[5]魏疆,禄邵英.miR-27在胃癌中的表达及其作用研究[J].陕西医学杂志,2015,44(5):620
[6]SUN Y,YANG X,LIU M,et al.B4GALT3 up-regulation by miR-27a contributes to the oncogenic activity in human cervical cancer cells[J].Cancer Lett,2016,375(2):284
[7]LIU J,MAO Y,ZHANG D,et al.MiR-143 inhibits tumor cell proliferation and invasion by targeting STAT3 in esophageal squamous cell carcinoma[J].Cancer Lett,2016,373(1):97
[8]TANG W,ZHU J,SU S,et al.MiR-27 as a prognostic marker for breast cancer progression and patient survival[J].PLo S One,2012,7(12):e51702
[9]MCALLISTER SD,MURASE R,CHRISTIAN RT,et al.Pathways mediating the effects of cannabidiol on the reduction of breast cancer cell proliferation,invasion,and metastasis[J].Breast Cancer Res Treat,2011,129(1):37
[10]杨浚沨,王龙强,李海,等.乳腺癌循环miRNA生物标志物的筛选及验证[J].中国普通外科杂志,2015,24(5):696
[11]QIN W,ZHANG K,CLARKE K,et al.Methylation and miRNA effects of resveratrol on mammary tumors vs.normal tissue[J].Nutr Cancer,2014,66(2):270
[12]CELI-TERRASSA T,LIU DD,CHOUDHURY A,et al.Normal and cancerous mammary stem cells evade interferon-induced constraint through the miR-199a-LCOR axis[J].Nat Cell Biol,2017,19(6):711
[13]PENG H,WANG X,ZHANG P,et al.miR-27a promotes cell proliferation and metastasis in renal cell carcinoma[J].Int J Clin Exp Pathol,2015,8(2):2259
[14]ZHOU L,LIANG X,ZHANG L,et al.MiR-27a-3p functions as an oncogene in gastric cancer by targeting BTG2[J].Oncotarget,2016,7(32):51943
[15]WANG YL,GONG WG,YUAN QL.Effects of miR-27a upregulation on thyroid cancer cells migration,invasion,and angiogenesis[J].Genet Mol Res,2016,15(4):38
[16]WANG T,XU Z.miR-27 promotes osteoblast differentiation by modulating Wnt signaling[J].Biochem Biophys Res Commun,2010,402(2):186
[17]顾文元.利拉鲁肽对人乳腺癌MCF-7细胞增殖、凋亡的影响及其机制研究[D].天津:天津医科大学,2016.
[18]ZHOU J,LI K,GU Y,et al.Transcriptional up-regulation of Rho E by hypoxia-inducible factor(HIF)-1 promotes epithelial to mesenchymal transition of gastric cancer cells during hypoxia[J].Biochem Biophys Res Commun,2011,415(2):348
[19]CHOI BD,JEONG SJ,WANG G,et al.Secretory leukocyte protease inhibitor is associated with MMP-2 and MMP-9 to promote migration and invasion in SNU638 gastric cancer cells[J].Int J Mol Med,2011,28(4):527
[20]BURLAKA AP,GANUSEVICH II,GAFUROV MR,et al.Stomach cancer:interconnection between the redox state,activity of MMP-2,MMP-9 and stage of tumor growth[J].Cancer Microenviron,2016,9(1):27