| Objectives: To observe the effect of rosiglitazone on the expression of LIMK1 and uPA in gastric cancer xenografts in nude mice,and the relationship between LIMK1 and uPA.Methods: To observe the morphology and structure of tumor tissues by HE stian,quantitative real-time PCR(qPCR)was used to detect the expression of LIMK1 and uPA gene in the tumor tissue of rosiglitazone group and normal saline group mRNA,The expression of LIMK1 and uPA protein in transplanted tumor tissues of each group were detected by Western blotting and immunohistochemistry(P-V method).Results: 1.Effect on the morphology on transplanted tumor(1)Effect of rosiglitazone on transplanted tumor : the corresponding rosiglitazone group compared with the normal saline group have a relatively large atypia,cells arranged orderly chaos,more intercellular substance,deeply nuclear staining,more than Caryoplasm,.Among them,In the normal saline group LIMK1 overexpression group was the most obvious,specificity of LIMK1 silence and the rosiglitazone intragastric administration group was the smallest.(2)The effect of mRNA on the expression of LIMK1 and uPA gene :The expression of LIMK1 and uPA gene in tumor tissue(2.4.6.8.10)decreased after rosiglitazone treatment,and the difference was statistically significant between the mRNA and the normal saline group(P<0.05).(3)The expression of limk and uPA protein in tumor tissues was detected by Western blotting:The expression of LIMK1 and uPA protein in the tumor tissues after rosiglitazone treatment decreased,and the difference was statistically significant compared with that of the normal saline group(P < 0.01).(4)The positive rates of LIMK1 and uPA protein were detected by immunohistochemistry : LIMK1,uPA positive tissue showed brown.The positive rate of LIMK1 and uPA rosiglitazone treated tissues(2.4.6.8.10)compared with normal saline group(1.3.5.7.9)was decline,The difference was statistically significant(P<0.05).2.Relationship between limk expression and uPA expression in transplanted tumor tissues of each group(In order to avoid the effect of rosiglitazone on the results,we selected the experimental data of the normal saline group).(1)qPCR detection of transplanted tumor:The expression of LIMK1 gene mRNA,LIMK1 overexpression group was increased,while LIMK1 decreased the expression of both silent group,the two group compare with MGC803 group had statistically significant difference(P<0.05),over expression vector group and silence vector group compared with MGC803 group,no significant difference(P>0.05).mRNA expression of uPA gene:LIMK1 overexpression group was increased,while LIMK1 decreased the expression of both silent group,the two group compare with MGC803 group had statistically significant difference(P<0.05),over-expression vector group and silence vector group compared with MGC803 group,no significant difference(P> 0.05).The relative expression of LIMK1 gene mRNA and the relative expression of uPA gene mRNA in transplanted tumor of each group were related to r = 0.618.It indicated that LIMK1 was positively related to uPA expression.(2)Detection of the expression of transplanted tumor protein by Western blotting:The expression of LIMK1 protein: LIMK1 protein of the LIMK1 overexpression group express more than the normal MGC803 cell group,there were statistically significant differences(P<0.01),while the expression of LIMK1 silencing group express more than MGC803 Cells group,the difference was statistically significant(P<0.01).Over expression vector group silent and empty vector group and MGC803 expression group had no statistically significant difference(P>0.05);the expression of uPA protein: the uPA protein of the LIMK1 overexpression group express more than the normal MGC803 cell group,there were statistically significant differences(P<0.01),while the expression of LIMK1 silencing group express more than MGC803 Cells group,the difference was statistically significant(P<0.01).Over expression vector group silent and empty vector group and MGC803 expression group had no statistically significant difference(P>0.05).The correlation between the positive rate of LIMK1 in LIMK1 over expression group and the positive rate of uPA was r = 0.388,The correlation between the positive rate of LIMK1 and the positive rate of uPA in the LIMK1 silencing group was r = 0.572.two results showed that the positive rate of two protein was positively correlated.(3)Immunohistochemical positive rate of tumor protein detection: the positive rate of LIMK1 expression: LIMK1 positive expression rate of MGC803 was higher than that of the normal group,the difference was significant(P<0.05),limk silencing group positive rate than normal MGC803 group were lower,the differences were statistically significant(P<0.05),and the expression vector group and silence vector group with normal MGC803 expression group had no statistically significant difference(P>0.05)the positive rate of.Upa expression: LIMK1 positive expression rate of MGC803 was higher than that of the normal group,the difference was significant(P<0.05),LIMK1 silencing group positive rate than normal MGC803 group were lower,the difference was statistically significant(P<0.05),and the expression vector group and silence vector group with normal MGC803 expression group had no statistically significant difference(P>0.05).The correlation between LIMK1 positive rate and uPA positive rate was r = 0.783 in each group of transplanted tumor,The positive rate of two protein expression was positively correlated.Conclusion:1.Rosiglitazone may inhibit MGC-803 cell growth in nude mice.2.LIMK1 may be rosiglitazone inhibits proliferation of human gastric cancer MGC-803 cells,target migration and invasion.3.The relationship between LIMK1 and uPA in gastric carcinoma was closely correlated with uPA.The invasion,migration and proliferation of tumor with high expression of.Limk1 may be related to the expression of the tumor... |
PDF Full Text Request