连接黏附分子C在乳腺癌脑转移组织中的表达及其对细胞侵袭性的影响  

Expression of binding molecule C in breast cancer brain metastases and its effect on cell invasion

作　　者：李世伟 张善义[1] Li Shiwei;Zhang Shanyi(Department of Neurosurgery,Sun Yat-sen Memorial Hospital,Guangzhou 510120,China)

机构地区：[1]中山大学孙逸仙纪念医院神经外科,广州510120

出　　处：《中华实验外科杂志》2023年第2期274-278,共5页Chinese Journal of Experimental Surgery

基　　金：广东省自然科学基金资助项目(2018A0303130193、2022A1515012393);广东省医学科研基金面上项目(A2016072);逸仙启航基金面上项目(YXQH201803)。

摘　　要：目的探讨连接黏附分子C(JAM-C)在乳腺癌脑转移瘤组织及乳腺癌细胞中的表达水平及其对肿瘤细胞迁徙、侵袭的影响。方法所需的人乳腺癌细胞株MDA-231、BT-549、SKBR3、T47D、MCF-7购自美国典型菌种保藏中心(ATCC)。人乳腺癌脑转移细胞株MDA-231-BM由美国国立癌症研究所(NCI)Patricia S.Steeg教授赠送。免疫组织化学检测JAMs、整联蛋白β3(ITGβ3)蛋白表达水平。通过实时荧光定量聚合酶链反应(qPCR)、蛋白质印迹法(Western blot)检测不同侵袭性人乳腺癌细胞中JAM-C基因mRAN及蛋白表达水平。小干扰RNA(siRNA)-JAM-C转染人乳腺癌细胞系MCF-7,Western blot检测细胞中JAM-C表达变化,Transwell小室法检测肿瘤细胞迁移和侵袭能力。两组间比较采用Student’s-t检验。结果免疫组化检测发现,JAM-C在脑转移乳腺癌组织中表达降低(脑转移乳腺癌比原位乳腺癌组织为2.36±0.82比3.12±0.93,t=3.967,P<0.05);ITGβ3在乳腺癌脑转移瘤组织中的表达高于原位乳腺癌(脑转移比原位乳腺癌组织为4.53±0.77比1.12±0.93,t=2.908,P<0.05)。实时荧光定量PCR发现,JAM-C基因mRNA水平在MDA231-BM(脑转移型)中低于低侵袭性乳腺癌细胞株MCF-7(脑转移株比乳腺癌株为1.53±0.35比3.18±0.76,P<0.05);Western blot结果显示,JAM-C蛋白水平亦显著低于后者(脑转移株比乳腺癌株为1.23±0.44比3.59±0.92,P<0.05)。划痕实验结果显示,敲除JAM-C实验组在划痕24 h后相对于对照组迁移距离更长[划痕愈合比(53.61±4.98)%比(32.55±2.63)%,P<0.05]。Transwell迁移、侵袭实验结果表明,沉默JAM-C可使实验组肿瘤细胞迁移能力显著高于对照组[(589±9)个比(269±14)个,t=41,P<0.01],侵袭能力显著高于对照组[(468.00±21.33)个比(212.00±35.04)个,t=32.45,P<0.05]。结论连接黏附分子C乳腺癌肿瘤细胞侵袭能力相关,具有抑制乳腺癌细胞迁移、侵袭的作用。Objective To investigate the expression level of JAM-C in breast cancer brain metastases(BMs)and breast cancer cells and its effect on tumor cell migration and invasion.Methods The human breast cancer cell lines MD-231,BT-549,SKBR3,T47D and MCF-7,and human breast cancer BMs cell line MDA-231-BM were applied.US Biomax Inc.(USA)normal breast tissue,primary breast cancer,invasive breast cancer and breast cancer BMs tissue chips were used to detect protein expression of JAMs.Quantitative real-time polymerase chain reaction(qPCR)and Western blotting were used to detect the expression levels of JAM-C mRAN and protein in different invasive breast cancer cells.Small interfering RNA(siRNA)-JAM-C was transfected into human breast cancer cell line MCF-7,and the expression of JAM-C in the cells was detected by Western blotting.The migration and invasion ability of tumor cells was detected by Transwell assay.Student’s t test was used for comparison between the two groups.Results The expression of JAM-C was decreased in BMS compared with in situ breast cancer(2.36±0.82 vs.3.12±0.93,t=3.967,P<0.05).The expression of ITGβ3 in BMS of breast cancer was higher than that in situ breast cancer(4.53±0.77 vs.1.12±0.93,t=2.908,P<0.05).Real-time quantitative PCR showed that the mRNA level of JAM-C gene in MDA231-BM(BMS)was lower than that in less invasive breast cancer cell line MCF-7(BMS vs.breast cancer strains were 1.53±0.35 vs.3.18±0.76,P<0.05).Western blotting results showed that the JAM-C protein level was also significantly lower than that of the latter(1.23±0.44 vs.3.59±0.92 in BMS vs.breast cancer strains,P<0.05).Scratch test results showed that the migration distance of the JAM-C knockout group was longer than that of the control group 24 h after scratch[scratch healing ratio(53.61±4.98)%vs.(32.55±2.63)%,P<0.05].Transwell migration and invasion experiments showed that the migration ability of tumor cells in the experimental group was significantly higher than that in the control group[(589±9)vs.(269±14),t=41,P<0.

关 键 词：连接黏附分子C 乳腺癌 脑转移 

分 类 号：R737.9[医药卫生—肿瘤]