张卫芳，孙　燕，张伟杰，朱　换，王留兴#

1)郑州大学第一附属医院肿瘤科一病区 郑州 450052　2)中国医学科学院肿瘤医院肿瘤内科 北京 100021



TM4SF1在人乳腺癌细胞MCF-7和MDA-MB-231中的表达及其对细胞增殖、迁移能力的影响

张卫芳1)，孙燕2)，张伟杰1)，朱换1)，王留兴1)#

1)郑州大学第一附属医院肿瘤科一病区 郑州 4500522)中国医学科学院肿瘤医院肿瘤内科 北京 100021

关键词TM4SF1；乳腺癌；转染；细胞增殖；迁移

摘要目的：探讨TM4SF1在人乳腺癌细胞MCF-7和MDA-MB-231中的表达情况以及靶向抑制TM4SF1基因后2种细胞的增殖、迁移能力的变化。方法：采用RT-PCR检测MCF-7和MDA-MB-231细胞中TM4SF1 mRNA表达情况，以正常乳腺上皮细胞MCF-10A的TM4SF1 mRNA表达作为参照；应用TM4SF1 siRNA瞬时转染MCF-7、MDA-MB-231细胞，应用Western blot方法分析转染效果和TM4SF1蛋白表达情况；CCK-8法检测沉默TM4SF1前后2种细胞增殖能力的变化；划痕实验检测细胞的迁移能力。结果：MCF-7细胞中TM4SF1 mRNA表达量为(1.159±0.218)，MDA-MB-231细胞中表达量为(1.396±0.199)，均高于MCF-10A细胞中的表达量(0.016±0.004)(P均<0.05)；且恶性程度较高的细胞MDA-MB-231高于低度恶性的MCF-7细胞(P<0.05)。特异性转染TM4SF1 siRNA后，该基因表达被抑制，同时MCF-7和MDA-MB-231细胞的体外迁移能力下降(P均<0.05)，而细胞的增殖能力变化差异无统计学意义(P>0.05)。结论：TM4SF1在人乳腺癌细胞MCF-7和MDA-MB-231中高表达，并增强乳腺癌细胞的迁移能力。

Expression of TM4SF1 and its effect on proliferation and migration of human breast cancer cells MCF-7 and MDA-MB-231

ZHANGWeifang1),SUNYan2),ZHANGWeijie1),ZHUHuan1),WANGLiuxing1)

1)WardOneofOncology,theFirstAffiliatedHospital,ZhengzhouUniversity,Zhengzhou450052 2)DepartmentofOncology,CancerHospital,ChineseAcademyofMedicalSciences,Beijing100021

Key wordsTM4SF1;breast cancer;transfection;cell proliferation;migration

AbstractAim: To explore the expression of TM4SF1 mRNA in MCF-7 and MDA-MB-231 cells, and investigate its effect on the proliferation and migration when TM4SF1 was silenced.Methods: The expression of TM4SF1 mRNA in MCF-7 and MDA-MB-231 cells was determined by RT-PCR, and the results were compared with that of MCF-10A cells. RNA interference method was applied to transiently transfect siRNA targeting at TM4SF1 into MCF-7 and MDA-MB-231 cells, the transfection efficiency and the protein expression of TM4SF1 in MCF-7 and MDA-MB-231 cells were analyzed by Western blot, CCK-8 was used to measure the difference of cell proliferation when TM4SF1 was silenced,and the mobility was determined by the scratch test.Results: The expression of TM4SF1 mRNA in MCF-7 cells (1.159±0.218) and in MDA-MB-231 cells (1.396±0.199) were both significantly higher than the expression in MCF-10A cells (0.016±0.004)(P<0.05), and the expression of TM4SF1 mRNA in MDA-MB-231 cells was higher than that in MCF-7 cells (P<0.05). After the TM4SF1 siRNA was transfected into the cells，the TM4SF1 mRNA expression were significantly inhibited, while the capability of migration of the two cells were decreased(P<0.05), and the difference of cell proliferation had no significance(P>0.05).Conclusion: TM4SF1 is highly expressed in MCF-7 and MDA-MB-231 cells, and enhances the migration of the cells.

乳腺癌是女性最常见的肿瘤[1]。国内外研究[2]表明，TM4SF1与恶性肿瘤的血管形成密切相关，并可促进多种肿瘤的侵袭与转移。TM4SF1是四次跨膜蛋白L6超家族成员之一，在肺癌[3]、胰腺癌[4]、前列腺癌[5]等有异常高表达，在正常的血管内皮细胞呈低表达[6]。该研究通过检测人乳腺癌细胞MCF-7和MDA-MB-231中TM4SF1表达情况，同时瞬时转染这2种细胞，检测细胞的增殖和迁移能力的变化，探讨TM4SF1与乳腺癌的关系，为乳腺癌的临床治疗提供一个新的靶点。

1材料与方法

1.1细胞株及主要材料MCF-7和MDA-MB-231细胞株由郑州大学肿瘤干细胞实验室提供，MCF-10A细胞由郑州大学第一附属医院重点实验室惠赠；……