Screening And Characterization Of Gastric Carcinoma Subcell Line With Higher Invasive Potential

Objective To better understand the mechanism(s) gastric cancer invasion and metastasis, a Transwell chamber was used to screen a gastric carcinoma subcell line with higher invasive potential and its biologic characters were studyed.Methods Transwell chamber was used for the selection of tumor subline. Morphology of the cell lines was observed with HE staining;Change of E-cadherin and TIMP-1 protein level was measured by western blot; Then Transwell was used to detect the difference of migration between MKN-28 and MKN-28S10 cells; SABC immunohistochemical method was employed for detecting the expressions of NM23-H1 protein and the immunohistochemical result was given situated and quantitative analysis by image analysis; Flow cytometry (FCM) was employed to analyze cell cycle;Growth curves of cell lines were figured.Result A gastric carcinoma cell subcell line (named MKN-28S10) was established from its parent cell line MKN-28 with higher invasive potential.MKN-28S10 showed essentially the same morphous as MKN-28, The tumor appeared polygon,disparity size,bigger karyoplasmic ratio, round or ellipse of nucleus; And the expression of E-cadherin,TIMP-1 decreased significantly in the screened subcell line. Compared with MKN-28 (relative gray scale value 0.90±0.04), the expression level of E-cadherin in MKN-28S10 (relative gray scale value 0.42±0.03) was decreased significantly (P<0.05),while the expression of TIMP-1 protein was obviously decreased in MKN-28S10 ( relative gray scale being 0.52±0..06)than that in MKN-28(0.89±0.05)(P<0.05); The expression of NM23-H1 in MKN-28S10 was significantly lower than that in MKN-28 (P<0.05) ,the mean optical density (MOD) of NM23-H1 was significantly decreased in MKN-28S10 compared with MKN-28(P<0.05);Compared with MKN-28(61.75±2.06 per vision), the migrative ability of passing through the membrane millipore(100.25±0.50 per vision)was obviously increased in MKN-28S10 (P<0.05); Flow cytometry showed that the percentage was 66.7% of the MKN-28S10 cells in the S phase,a higher percentage than that of the MKN-28 cells, moreover, the proliferating index(PI) of MKN-28S10(74.6%)was higher than that of MKN-28(64.8%);As compared with MKN-28,the growth rate of MKN-28S10 was increased obviously.Conclusions1. Gastric carcinoma subcell line with higher invasive potential was screened2. Different invasive capacities between the parent (MKN-28) and the subline cell (MKN-28S10) are related to the expression levels of E-cadherin,TIMP-1, NM23-H1 and the migratory ability3. MKN-28S10 cell has more powerful proliferation than that of its mother line MKN-28...