CAJ | 학술논문

目的研究糖蛋白丝甘蛋白聚糖(SRGN)的表达与乳腺癌细胞侵袭转移的关系,及小分子RNA在SRGN表达调控中的作用.方法应用实时定量PCR和Western blot检测SRGN在侵袭转移增强的乳腺癌MCF-7/5-Fu细胞与亲本转移能力弱的MCF-7细胞中的表达差异,利用siRNA干扰技术结合体外Transwell实验检测SRGN对乳腺癌细胞侵袭转移能力的影响.运用生物信息学软件预测可靶向SRGN的miRNA,并对接乳腺癌细胞MCF-7 vs MCF-7/5-Fu的miRNA差异表达芯片数据,同时利用miRNA定量PCR确定候选miRNA的表达差异.通过转染miRNA模拟物结合western blot实验验证候选miR-NA对SRGN靶向调节作用,最后通过Transwell实验检测候选miRNA对肿瘤细胞侵袭转移的影响.结果 SRGN在侵袭转移增强的MCF-7/5-Fu细胞中表达明显增高,干扰SRGN可抑制肿瘤细胞的侵袭转移.此外,miR181b/c在MCF-7/5-Fu细胞中表达明显降低与SRGN表达呈负相关,并且miR181 b/c可靶向抑制SRGN表达,进而抑制肿瘤细胞的侵袭转移.结论 miR181 b/c可靶向抑制SRGN表达,抑制乳腺癌细胞的侵袭转移.
목적 연구당단백사감단백취당(SRGN)적표체여유선암세포침습전이적관계,급소분자RNA재SRGN표체조공중적작용.방법 응용실시정량PCR화Western blot검측SRGN재침습전이증강적유선암MCF-7/5-Fu세포여친본전이능력약적MCF-7세포중적표체차이,이용siRNA간우기술결합체외Transwell실험검측SRGN대유선암세포침습전이능력적영향.운용생물신식학연건예측가파향SRGN적miRNA,병대접유선암세포MCF-7 vs MCF-7/5-Fu적miRNA차이표체심편수거,동시이용miRNA정량PCR학정후선miRNA적표체차이.통과전염miRNA모의물결합western blot실험험증후선miR-NA대SRGN파향조절작용,최후통과Transwell실험검측후선miRNA대종류세포침습전이적영향.결과 SRGN재침습전이증강적MCF-7/5-Fu세포중표체명현증고,간우SRGN가억제종류세포적침습전이.차외,miR181b/c재MCF-7/5-Fu세포중표체명현강저여SRGN표체정부상관,병차miR181 b/c가파향억제SRGN표체,진이억제종류세포적침습전이.결론 miR181 b/c가파향억제SRGN표체,억제유선암세포적침습전이.
Objective To investigate the relationship of glycoprotein serglycin (SRGN) expression with invasion and metastasis of breast cancer cells,and the role of microRNA in the regulation of SRGN expression.Methods Real-time quantitative polymer ase chain reaction (PCR) and Western blot were used to detect the differences in SRGN expression between higher metastasis Michigan cancer foundation-7 (MCF-7)/5-Fu breast cancer cell lines and weaker metastasis MCF-7 cell line.The siRNA interference experiment and in vitro Transwell experiment were used to detect effect of SRGN on the ability of invasion and metastasis of breast cancer cells.Bioinformatics software was used to predict miRNAs targeting SRGN,and integrated microRNA differentially expressed chip data between breast cancer cell MCF-7 versus MCF-7/5-Fu.The miRNA quantitative PCR was used to determine the differences of candi date miRNA expression.After transfection of microRNA minics,Western blot was used to test candidate microRNA target SRGN.Transwell experiment was used to test the effects of candidate microRNAs on tumor cell invasion and metastasis.Results SRGN was increased significantly in MCF-7/5-Fu cells,and the invasion and metastasis of tumor cells were inhibited when SRGN was interfered.In addition,miR181 b/c expressed in MCF-7/5-Fu cells was reduced significantly,negatively correlated with SRGN expression,and targeted SRGN expression.It inhibited invasion and metastasis of tumor cells.Conclusions MicroRNA181b/c inhibits metastasis of breast cancer by targeting SRGN.