摘要
目的观察抗肝肠钙粘连蛋白（CDHl7）单克隆抗体Lie5对肝癌细胞生物学行为的影响。方法Westernblot和实时定量聚合酶链反应（PCR）检测细胞株MHCC97H、MHCC97L、PLC／PRF／5及MIHA中CDHl7的表达。噻唑蓝（MTr）比色法、细胞划痕法、Transwell法及平板克隆法检测Lie5对肝癌细胞生物学行为的影响。结某CDHl7仅在细胞株MHCC97H、MHCC97L中表达，Lic5可结合肝癌细胞表面的CDHl7，并抑制CDHl7表达。Lic550mg／L组、100mg／L组、小鼠IgG组4d细胞增殖抑制率在MHCC97H为26．1％、43．6％、6．4％，MHCC97L为26．O％、40．7％、7．7％；Lic5100mg／L组、小鼠IgG组、磷酸盐缓冲液（PBS）组48h细胞迁移抑制率在MHCC97H为36．7％、8．4％、5．6％，MHCC97L为42．3％、10．2％、7．4％（P〈0．05）；穿膜细胞数在MHCC97H为（39．20±9．56）、（106．50±7．56）、（96．60±3．02）个，MHCC97L为（26．00±8．61）、（86．00±10．26）、（90．40±12．04）（P〈0．05）；克隆形成数在MHCC97H为（59．30±11．68）、（141．70±19．40）、（150．30±14．64）。MHCC97L为（57．20±10．21）、（132．50±9．07）、（121．70±11．93）（P〈0．01）。Lic5对PLC／PRFl5及MIHA细胞的生物学行为无明显影响。结论单克隆抗体Lie5能够下调肝癌细胞CDH17表达，抑制肝癌细胞的增殖、迁移、侵袭和克隆形成能力。
Objective To investigate the effect of monoclonal antibody against liver-intestine cadherin （CDH17） on the cell proliferation, migration, invasion and colony formation of hepatocellular carcinoma cell lines. Methods Hepatocellular carcinoma cell lines MHCC97H, MHCC97L, PLC/PRF/5 and MIHA were examined for CDH17 expression by Western blotting and quantitative real-time polymerase chain reaction （PCR）. The combination capacity between hepatocellular carcinoma cell lines and monoclonal antibody Lic5 was detected by the way of immunofluorescence staining. The cell lines were treated with LicS, PBS and mouse IgG respectively. Methyl thiazol tetrazolium （MTT） assay, wound healing assay, Transwell invasion assay and colony formation assay were used to study the changes in cell proliferation, migration, invasion and colony formation of hepatocellular carcinoma cell lines. Results High expression level of CDH17 was detected in MHCC97H and MHCC97L cell lines. CDH17 protein level was down-regulated but there was no significant effect on CDH17 mRNA after treatment with Lic5 in MHCC97H and MHCC97L. Cellular growth rate of MHCC97H in Lic5 （50 mg/L）, Lic5 （100 mg/L） and mouse IgG groups was decreased by 26. 1% , 43.6% and 6. 4% , and by 26. 0% , 40. 7% and 7. 7% in MHCC97L on the 4th day respectively （P 〈0. 05）. The inhibition rate of cell migration at 48 h was 36.7%, 8. 4% and 5.6% in Lic5 （100 mg/L）, mouse IgG and PBS groups in MHCC97H, and 42. 3% , 10. 2% and 7. 4% in MHCC97L respectively （ P 〈 0. 05 ）. The number of invasion cells was （ 39. 20 ± 9. 56 ）, （106. 50 ±7.56） and （96. 60±13.02） in MHCC97H, and （26.00 ±8.61）, （86.00±10.26） and （90.40 ± 12. 04） in MHCC97L in Lic5 （50 mg/L）, Lic5 （ 100 mg/L） and mouse IgG groups, respectively （P 〈 0. 05 ）. The number of colony formation was （ 59.30 ± 11.68 ）, （ 141.70 ± 19.40 ） and （150.30±14.64） inMHCC97H, and （57.20±10.21）, （132.50 ±9.07） and （121.70±11.93） in MHCC97L in L
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2011年第8期1295-1298,共4页
Chinese Journal of Experimental Surgery
关键词
肝肠钙粘连蛋白
单克隆抗体
癌
肝细胞
Liver-intestine cadherin
Monoclonal antibody
Carcinoma, hepatocellular
