Yu You, Han De-En, Liu Wei
Department of Pediatric Surgery, The Second Affiliated Hospital,Harbin Medical University, Harbin, Heilongjiang, 150086, PR China.
Ai Zheng. 2007 May;26(5):498-503.
BACKGROUND & OBJECTIVE: Metastasis suppressor gene KAI1 can inhibit the metastasis potential of many malignant tumors. This study was to explore the effect of KAI1 gene on the adhesion of hepatocellular carcinoma cell line MHCC97-H with high metastatic potential, and to find the possible mechanism.
The plasmid containing KAI1 gene was transfected into MHCC97-H cells. The growth status of the cells was observed. The changes of adhesion factors, soluble intercellular adhesive molecule-1 (sICAM-1) and E-cadherin, in the cells were detected by ELASA and Western blot. The character of cell adhesion was determined through plate colony formation test and cell adhesion test.
The morphology of MHCC97-H cells did not change markedly after transfection of KAI1 gene, but black granules in cytoplasm were increased. Four days after transfection, the expression of sICAM-1 and E-cadherin were decreased by 24.28% and 26.02%. The adhesion rate was decreased by 11.34% at 3 h after transfection, and by 24.00% at 4 h after transfection, but the cloning efficiency did not change much.
KAI1 gene could affect the growth pattern and proliferation of MHCC97-H cells, suppress sICAM-1 secretion and E-cadherin production, and inhibit adhesion of MHCC97-H cells.
转移抑制基因KAI1可抑制多种恶性肿瘤的转移潜能。本研究旨在探讨KAI1基因对高转移潜能肝癌细胞系MHCC97-H黏附的影响,并寻找可能的机制。
将含KAI1基因的质粒转染至MHCC97-H细胞。观察细胞生长状态。采用酶联免疫吸附测定法(ELASA)和蛋白质免疫印迹法检测细胞中黏附因子可溶性细胞间黏附分子-1(sICAM-1)和E-钙黏蛋白的变化。通过平板集落形成试验和细胞黏附试验确定细胞黏附特性。
转染KAI1基因后,MHCC97-H细胞形态无明显变化,但细胞质中黑色颗粒增多。转染后4天,sICAM-1和E-钙黏蛋白的表达分别下降了24.28%和26.02%。转染后3小时黏附率下降了11.34%,4小时下降了24.00%,但克隆效率变化不大。
KAI1基因可影响MHCC97-H细胞的生长模式和增殖,抑制sICAM-1分泌和E-钙黏蛋白产生,并抑制MHCC97-H细胞的黏附。