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目的探讨不同乳腺癌细胞系体外侵袭能力的差异并筛选其影响基因。方法以2种常见的乳腺癌细胞系MCF-7(低转移株)和MDA-MB-231(高转移株)为研究对象,通过细胞划痕实验、细胞黏附实验以及Transw ell侵袭实验分析2种乳腺癌细胞体外侵袭能力差异,实时荧光定量PCR筛选肿瘤侵袭转移相关基因在细胞间的表达差异并采用细胞免疫组化SP法进一步确认。结果 MDA-MB-231细胞24、48 h的平均迁移距离分别为(218.75±20.16)、(211.50±16.54)μm,明显高于MCF-7细胞的迁移距离[分别为(130±29.62)、(119.50±25.65)μm](P<0.01);MDA-MB-231细胞黏附能力明显高于MCF-7细胞;MDA-MB-231细胞24 h平均穿膜细胞数[(57.75±4.19)个]明显多于MCF-7细胞[(35.50±4.20)个](P<0.01);实时荧光定量PCR结果显示,与MCF-7细胞比较,MDA-MB-231细胞中细胞间黏附分子1(ICAM-1)及乳腺癌候选抑制蛋白1(BCSC-1)表达水平[分别为(0.09±0.01)、(0.15±0.03)]均明显降低(P<0.01),而基质金属蛋白酶14(MMP-14)表达水平(14.43±1.01)明显升高(P<0.01)。细胞免疫组化结果显示,MDA-MB-231细胞ICAM-1以及BCSC-1的表达水平明显低于M CF-7细胞,而M M P-14表达水平明显高于M CF-7细胞。结论乳腺癌细胞系M DA-M B-231的体外侵袭能力明显高于MCF-7细胞,细胞侵袭力可能与细胞内ICAM-1、BCSC-1低表达及MMP-14高表达有关。
Objective To investigate the difference of invasive ability of different breast cancer cell lines in vitro and to screen the genes involved. Methods Two common breast cancer cell lines, MCF-7 (low metastatic strain) and MDA-MB-231 (highly metastatic strain), were studied by cell scratch assay, cell adhesion assay and Transwell assay. Breast cancer cells in vitro invasive ability differences, real-time fluorescence quantitative PCR screening tumor invasion and metastasis-related genes in the cell differences in expression and further confirmed by immunohistochemical SP method. Results The average migration distances of MDA-MB-231 cells at 24 and 48 h were (218.75 ± 20.16) and (211.50 ± 16.54) μm respectively, which were significantly higher than those of MCF-7 cells [130 ± 29.62 and 119.50 ± 25.65) μm] (P <0.01). The adhesion capacity of MDA-MB-231 cells was significantly higher than that of MCF-7 cells (P <0.01). Compared with MCF-7 cells, the expression of intercellular adhesion molecule 1 (ICAM-1) in MDA-MB-231 cells was significantly higher than that in MCF-7 cells ) And BCSC-1 expression (0.09 ± 0.01, 0.15 ± 0.03, respectively) were significantly decreased (P <0.01), while the expression of matrix metalloproteinase-14 Level (14.43 ± 1.01) was significantly higher (P <0.01). The results of immunohistochemistry showed that the expression level of ICAM-1 and BCSC-1 in MDA-MB-231 cells was significantly lower than that in M CF-7 cells, while the expression level of M M P-14 was significantly higher than that in M CF-7 cells. Conclusion The invasiveness of breast cancer cell line M DA-M B-231 in MCF-7 cells was significantly higher than that of MCF-7 cells. The invasiveness of MCF-7 cells may be related to the low expression of ICAM-1 and BCSC-1 and the high expression of MMP-14.