CUG连接蛋白1在乳腺癌组织中的表达及其对细胞活力和侵袭能力的影响
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摘要
目的:探讨CUG连接蛋白1(CUGBP1)在乳腺癌组织中的表达,以及沉默CUGBP1基因对人乳腺癌MCF-7细胞活力和侵袭能力的影响。方法:选取2015年3月～2017年9月在郑州大学第二附属医院手术治疗的乳腺癌患者96例,免疫组化法检测乳腺癌和癌旁组织中CUGBP1蛋白表达,培养MCF-7细胞并分为CUGBP1干扰序列组、对照序列组和空白组,Western blot法检测各组细胞中CUGBP1、Twist、上皮型钙黏附蛋白(E-cadherin)和波形蛋白(vimentin)的蛋白表达,MTT法检测各组细胞活力,Transwell法检测细胞侵袭能力。结果:乳腺癌组织中CUGBP1阳性表达率高于癌旁组织(χ~2=28.900,P<0.001);乳腺癌组织中CUGBP1蛋白表达在TNM分期、组织学分级和淋巴结转移差异有统计学意义(P<0.05);CUGBP1干扰序列组细胞中CUGBP1、Twist和vimentin蛋白相对表达量均低于对照序列组和空白组,而E-cadherin蛋白相对表达量均高于对照序列组和空白组(P<0.05);CUGBP1干扰序列组24 h、48 h、72 h和96 h时细胞活力均低于对照序列组和空白组(P<0.05);CUGBP1干扰序列组侵袭细胞数少于对照序列组和空白组(P<0.05)。结论:乳腺癌组织中CUGBP1蛋白呈高表达。特异性沉默乳腺癌MCF-7细胞CUGBP1基因表达,可有效抑制细胞活力及侵袭能力,其机制可能与抑制上皮-间充质转化过程有关。
AIM: To investigate the expression of CUG-binding protein 1(CUGBP1) in breast cancer tissues, and to explore the effect of CUGBP1 gene silencing on the viability and invasion ability of human breast cancer MCF-7 cells. METHODS: A total of 96 cases of patients with breast cancer undergoing surgical treatment were selected in the Second Affiliated Hospital of Zhengzhou University from March 2015 to September 2017. Immunohistochemical staining was used to detect the protein expression of CUGBP1 in the breast cancer and adjacent tissues. MCF-7 cells were cultured and divided into CUGBP1 interference sequence group, control sequence group and blank group. Western blot was used to detect the protein expression of CUGBP1, Twist, E-cadherin and vimentin in the cells. The cell viability was measured by MTT assay. The cell invasion ability was detected by Transwell assay. RESULTS: The positive expression rate of CUGBP1 protein in the breast cancer tissues was higher than that in the adjacent tissues(χ~2=28.900, P<0.001). The differences of CUGBP1 protein expression in the breast cancer tissues among TNM staging, histological grading and lymph node metastasis were statistically significant(P<0.05). The relative protein expression levels of CUGBP1, Twist and vimentin in CUGBP1 interference sequence group were lower than those in control sequence group and blank group, while the relative protein expression of E-cadherin was higher than that in control sequence group and blank group(P<0.05). The cell viability at 24 h, 48 h, 72 h and 96 h in CUGBP1 interference sequence group was lower than that in control sequence group and blank group(P<0.05). The invasive cells in CUGBP1 interference sequence group were less than those in control sequence group and blank group(P<0.05). CONCLUSION: CUGBP1 protein is highly expressed in the breast cancer tissues. Specific silencing of CUGBP1 gene expression in breast cancer MCF-7 cells effectively inhibits the cell viability and invasiveness, and its mechanism may be related to inhibiting the process of epithelial-mesenchymal transition.
AIM: To investigate the expression of CUG-binding protein 1(CUGBP1) in breast cancer tissues, and to explore the effect of CUGBP1 gene silencing on the viability and invasion ability of human breast cancer MCF-7 cells. METHODS: A total of 96 cases of patients with breast cancer undergoing surgical treatment were selected in the Second Affiliated Hospital of Zhengzhou University from March 2015 to September 2017. Immunohistochemical staining was used to detect the protein expression of CUGBP1 in the breast cancer and adjacent tissues. MCF-7 cells were cultured and divided into CUGBP1 interference sequence group, control sequence group and blank group. Western blot was used to detect the protein expression of CUGBP1, Twist, E-cadherin and vimentin in the cells. The cell viability was measured by MTT assay. The cell invasion ability was detected by Transwell assay. RESULTS: The positive expression rate of CUGBP1 protein in the breast cancer tissues was higher than that in the adjacent tissues(χ~2=28.900, P<0.001). The differences of CUGBP1 protein expression in the breast cancer tissues among TNM staging, histological grading and lymph node metastasis were statistically significant(P<0.05). The relative protein expression levels of CUGBP1, Twist and vimentin in CUGBP1 interference sequence group were lower than those in control sequence group and blank group, while the relative protein expression of E-cadherin was higher than that in control sequence group and blank group(P<0.05). The cell viability at 24 h, 48 h, 72 h and 96 h in CUGBP1 interference sequence group was lower than that in control sequence group and blank group(P<0.05). The invasive cells in CUGBP1 interference sequence group were less than those in control sequence group and blank group(P<0.05). CONCLUSION: CUGBP1 protein is highly expressed in the breast cancer tissues. Specific silencing of CUGBP1 gene expression in breast cancer MCF-7 cells effectively inhibits the cell viability and invasiveness, and its mechanism may be related to inhibiting the process of epithelial-mesenchymal transition.
引文
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[12] Yang L,Zhang J,Chen J,et al.The expression of CUGBP1 after spinal cord injury in rats[J].Neurochem Res,2015,40(9):1966-1975.
[13] Xia L,Sun C,Li Q,et al.CELF1 is up-regulated in glioma and promotes glioma cell proliferation by suppression of CDKN1B[J].Int J Biol Sci,2015,11(11):1314-1324.
[14] Lu H,Yu Z,Liu S,et al.CUGBP1 promotes cell proli-feration and suppresses apoptosis via down-regulating C/EBPα in human non-small cell lung cancers[J].Med Oncol,2015,32(3):82-88.
[15] Gao C,Yu Z,Liu S,et al.Overexpression of CUGBP1 is associated with the progression of non-small cell lung can-cer[J].Tumour Biol,2015,36(6):4583-4589.
[16] Bednarczyk RB,Tuli NY,Hanly EK,et al.Macrophage inflammatory factors promote epithelial-mesenchymal transition in breast cancer[J].Oncotarget,2018,9(36):24272-24282.
[17] Xie X,Zheng X,Wang J,et al.Clinical significance of Twist,E-cadherin,and N-cadherin protein expression in endometrioid adenocarcinoma[J].J Cancer Res Ther,2017,13(5):817-822.
[2] 徐兵河,王树森,江泽飞,等.中国晚期乳腺癌维持治疗专家共识[J].中华医学杂志,2018,98(2):87-90.
[3] 段堃,李岩岩,赵艳,等.CUG连接蛋白1在膀胱癌中的表达及其对BIU-87细胞株增殖及侵袭能力的影响[J].实用医学杂志,2016,32(3):411- 414.
[4] Wang X,Jiao W,Zhao Y,et al.CUG-binding protein 1 (CUGBP1) expression and prognosis of brain metastases from non-small cell lung cancer[J].Thorac Cancer,2016,7(1):32-38.
[5] Kim JH,Kwon HY,Ryu DH,et al.Inhibition of CUG-binding protein 1 and activation of caspases are critically involved in piperazine derivative BK10007S induced apoptosis in hepatocellular carcinoma cells[J].PLoS One,2017,12(10):e0186490.
[6] 李树斌,张延新.结肠癌转移相关基因1及c-met蛋白在乳腺癌组织中的表达及临床意义[J].中国老年学杂志,2018,38(4):819-820.
[7] Jacklyn G,McGeechan K,Irwig L,et al.Trends in stage-specific breast cancer incidence in New South Wales,Australia:insights into the effects of 25 years of screening mammography[J].Breast Cancer Res Treat,2017,166(3):843-854.
[8] Wang J,Gao S,Wang Y,et al.Cancer incidence and mortality patterns in Luwan district of Shanghai during 2002-2011[J].Drug Discov Ther,2018,12(2):77-87.
[9] Jurkovicova D,Smolkova B,Magyerkova M,et al.Down-regulation of traditional oncomiRs in plasma of breast can-cer patients[J].Oncotarget,2017,8(44):77369-77384.
[10] Cui YX,Bradbury R,Flamini V,et al.MicroRNA-7 suppresses the homing and migration potential of human endothelial cells to highly metastatic human breast cancer cells[J].Br J Cancer,2017,117(1):89-101.
[11] 朱清海,王艳瑛,张漫丽,等.CUG连接蛋白1在肝癌中的表达及其对细胞增殖能力的影响[J].中华实验外科杂志,2014,31(1):159-161.
[12] Yang L,Zhang J,Chen J,et al.The expression of CUGBP1 after spinal cord injury in rats[J].Neurochem Res,2015,40(9):1966-1975.
[13] Xia L,Sun C,Li Q,et al.CELF1 is up-regulated in glioma and promotes glioma cell proliferation by suppression of CDKN1B[J].Int J Biol Sci,2015,11(11):1314-1324.
[14] Lu H,Yu Z,Liu S,et al.CUGBP1 promotes cell proli-feration and suppresses apoptosis via down-regulating C/EBPα in human non-small cell lung cancers[J].Med Oncol,2015,32(3):82-88.
[15] Gao C,Yu Z,Liu S,et al.Overexpression of CUGBP1 is associated with the progression of non-small cell lung can-cer[J].Tumour Biol,2015,36(6):4583-4589.
[16] Bednarczyk RB,Tuli NY,Hanly EK,et al.Macrophage inflammatory factors promote epithelial-mesenchymal transition in breast cancer[J].Oncotarget,2018,9(36):24272-24282.
[17] Xie X,Zheng X,Wang J,et al.Clinical significance of Twist,E-cadherin,and N-cadherin protein expression in endometrioid adenocarcinoma[J].J Cancer Res Ther,2017,13(5):817-822.