A Preliminary Study On Relationship Of Invasiveness And Multi-drug Resistance In Parent And Sub-cell Line Of Human Gastric Carcinoma Cells

Objective To explore mechanism and relationship of invasiveness and metastasis and multi-drug resistance in parent and sub-cell line of human gastric carcinoma cells(MKN-28).Methods The invasiveness was assessed by penetration through a matrigel-coated filter in transwell chamber; Protein expression level of MMP-9, MMP-2, P-gp, MRP1, ERK1/2 and p-ERK1/2 in MKN-28S10 and MKN-28 cell lines was assessed by Western blot; Protein expression level of MMP-9, MMP-2, P-gp, MRP1 in MKN-28S10 treated with various concentrations of U0126 was assessed by Western blot; SP immunocytochemical method was employed for detecting the expressions of MMP-9, MMP-2, P-gp, MRP1 protein; MTT assays were used to detect sensitivity of two cell lines by ADM, MMC and 5-Fu treatment.Result The mount of sub-cell line (named MKN-28S10) cells penetrating through matrigel-coated filter in transwell chamber was significantly increased compared with parent cells(named MKN-28) under light microscope (P<0.05); Protein expression level of MMP-9, MMP-2, P-gp, MRP1 significantly increased in MKN-28S10 cell line compared with MKN-28 cell line; SP immunocytochemical result also show the expression of MMP-9, MMP-2, P-gp, MRP1 significantly increased in MKN-28S10 cell line compared with MKN-28 cell line; Protein expression level of p-ERK1/2 was significantly decreased in MKN-28S10 cell line compared with MKN-28 cell line (P<0.05), but not of ERK1/2 (P>0.05); The sensitivities in both cell lines were significantly different for ADM, MMC and 5-Fu (P<0.05); U0126, an inhibitor of MAPK/extracellular signal regulated kinase (ERK), led to significant dose-dependent (1-10μM) inhibition of expression of MMP-9, MMP-2, P-gp, MRP1 in MKN-28S10 cell line.Conclusions1. The invasive and metastatic potential in sub-cell line (named MKN-28S10) is significantly increased compared with parent cell line (named MKN-28);2. The multi-drug resistance in sub-cell line (named MKN-28S10) is significantly increased compared with parent cell line (named MKN-28);3. Alteration in human gastric tumor cells'invasiveness and metastasis can effect on its multi-drug resistance;4. An ERK1/2-dependent signaling pathway plays an important effect on regulating the pression of MMP-9, MMP-2, P-gp and MRP1.