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微小RNA-194对基底样乳腺癌细胞HCC1937增殖、迁移侵袭及上皮-间充质转化的影响

Effects of microRNA-194 on proliferation, migration, invasion and epithelial-mesenchymal transition of basal-like breast cancer cell line HCC1937

摘要:

目的 观察微小RNA-194(miR-194)对人基底样乳腺癌细胞(HCC1937)增殖、迁移侵袭能力及上皮-间充质转化(E MT)的影响.方法 利用慢病毒作载体(MOI=50)携带miR-194感染基底样乳腺癌细胞HCC1937为实验组,携带CON157慢病毒感染乳腺癌细胞HCC1937为阴性对照组,未做处理的HCC1937细胞为空白对照组.细胞计数试剂盒(CCK-8)法与Transwell小室实验分别检测HCC1937细胞增殖、迁移侵袭能力;Western blot方法检测EMT相关蛋白E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)的表达.结果 实验组细胞增殖能力为(44.81±0.11)%,显著低于阴性对照组为(104.29 ±0.13)%,空白对照组为(100±0)%(P <0.01);实验组细胞迁移及侵袭力显著低于阴性对照组及空白对照组(P<0.05);E-cadherin蛋白:实验组(0.175 ±0.053)显著低于阴性对照组(0.516 ±0.032)、空白对照组(0.433±0.047,P<0.05);N-cadherin蛋白表达实验组(0.342±0.012)显著低于阴性对照组(0.137 ±0.007)、空白对照组(0.142 ±0.021,P<0.01).结论 miR-194能抑制基底细胞样乳腺癌细胞HCC1937增殖、迁移侵袭能力及EMT.

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abstracts:

Objective To explore the role of microRNA-194 (miR-194) in proliferation,migration,invasion and the epithelial-mesenchymal transition (EMT) of human basal-like breast cancer cell line HCC1937.Methods Three groups were set up:experimental group [HCC1937 with lentivirus-miR-194-enhanced green fluorescent protein (EGFP),MOI =50],negative control group (HCC1937with lentivirus-CON157-EGFP),and blank control group (HCC1937 without lentivirus).The miR-194 expression was detected by real-time quantitative reverse transcriptase-polymerase chain reaction (RT-qPCR).Cell growth ability of HCC1937 was determined by cell counting kit-8 (CCK-8) assay.Cell migration and invasion ability of HCC1937 were mearsured by a Transwell assay,and the protein of epithelial-mesenchymal transition phenotype was measured by Western blotting.Results The expression of miR-194 in experimental group (432.93 ± 0.01) was higher than in negative control group (0.90 ±0.02) and blank control group (1) (P <0.01) The cells in experimental group grew more slowly [(44.81 ± 0.11)%] than negative control group [(104.29 ± 0.13)%] and blank control group [(100 ±0) %,P <0.01].The number of migration and invasion was both less in experimental group than in negative control group and blank control group (P < 0.05).Western blotting revealed that the expression of E-cadherin in experimental group (0.175 ± 0.053) was significantly decreased as compared with negative control group (0.516 ± 0.032) and blank control group (0.433 ± 0.047,P < 0.05),and the expression of N-cadherin in experimental group (0.342 ± 0.012) was significantly increased as compared with negative control group (0.137 ±0.007) and blank control group (0.142 ±0.021) (P <0.05).Conclusion miR-194 can inhibit proliferation,migration and invasion of basal-like breast cancer cells (HCC1937),and also affect EMT process in HCC1937 cells.

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作者: 孔吉霖 [1] 吕志栋 [2] 姜丹丹 [2] 柳晓义 [2] 刘相萍 [3] 孙苏园 [4] 辛海娜 [1] 李福年 [2]
期刊: 《中华实验外科杂志》2015年32卷5期 969-972页 ISTICPKU
栏目名称: 乳腺外科
DOI: 10.3760/cma.j.issn.1001-9030.2015.05.007
发布时间: 2015-06-04
基金项目:
国家自然科学基金青年科学基金资助项目 山东省自然科学基金资助项目
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