Ⅲ型胶原α1与乳腺癌细胞恶性进展相关性的实验研究

Experimental study on the correlation between typeⅢcollagenα1 and malignant progression of breast cancer cells

目的探讨Ⅲ型胶原α1(COL3A1)对乳腺癌(BRCA)细胞增殖、迁移、侵袭和血管生成的影响。方法GEPIA数据库用于分析COL3A1在BRCA组织和正常组织中的差异表达;定量逆转录聚合酶链式反应(qRT-PCR)用于检测正常乳腺上皮细胞(MCF-10A)和BRCA细胞中COL3A1的mRNA表达水平;CCK-8法、EDU实验、细胞划痕实验和Transwell实验和血管形成实验分别用于检测BRCA细胞的增殖、迁移、侵袭和血管生成;Western blot用于检测BRCA细胞中COL3A1和VEGFA蛋白的表达。在体内,构建裸鼠异种移植瘤模型。结果GEPIA数据库分析显示,与正常组织相比,COL3A1在BRCA组织中高表达(P<0.05)。在体外,与sh-NC相比,COL3A1敲除显著抑制了MCF-7和MDA-MB-231细胞的增殖、迁移、侵袭和血管生成(P<0.01)。与sh-NC相比,COL3A1敲除明显下调了VEGFA的蛋白表达(P<0.01)。在体内,与sh-NC相比,COL3A1敲除抑制了异种移植瘤的体内生长(P<0.01)。结论COL3A1在BRCA中高表达,COL3A1敲除可抑制BRCA细胞的增殖、迁移、侵袭和血管生成,可能是治疗BRCA新的生物标志物。

Objective To explore the effects of collagen typeⅢα1(COL3A1)on proliferation,migration,invasion and angiogenesis of breast cancer(BRCA)cells.Methods The GEPIA database was used to analyze the differential expression of COL3A1 in BRCA tissues and normal tissues.Quantitative reverse transcription polymerase chain reaction(qRT-PCR)was used to detect the mRNA expression level of COL3A1 in normal breast epithelial cell(MCF-10A)and BRCA cells.CCK-8 assay,EDU assay,cell scratch assay,Transwell assay and Tube formation assay were used to detect the proliferation,migration,invasion and angiogenesis of BRCA cells,respectively.Western blot was used to detect the expression of COL3A1 and VEGFA proteins in BRCA cells.In vivo,the xenograft tumor model in nude mice was established.Results GEPIA database analysis showed that COL3A1 was high expressed in BRCA tissues compared to normal tissues(P<0.05).In vitro,compared with sh-NC,COL3A1 knockout significantly inhibited the proliferation,migration,invasion and angiogenesis of MCF-7 and MDA-MB-231 cells(P<0.01).Compared with sh-NC,COL3A1 knockout significantly down-regulated the protein expression of VEGFA(P<0.01).In vivo,COL3A1 knockout inhibited the growth of xenograft tumors compared with sh-NC(P<0.01).Conclusion COL3A1 is highly expressed in BRCA,and the deletion of COL3A1 inhibites the proliferation,migration,invasion and angiogenesis of BRCA cells,which might be a new biomarker for the treatment of BRCA.