ABSTRACT
Objective To investigate the influence of sorafenib on the expressions of B7-H3 and B7-H4 proteins in different human hepatocellular carcinoma cell lines,including HepG2,Hep3B,BEL-7402,BEL-7404,BEL-7405,QGY-7701,QGY-7703,SMMC-7721,MHCC97H,MHCC97L,HCCLM3 and HCCLM6.Methods Western blotting and MTT assay were used to check the influence of sorafenib on the expressions of B7-H3 and B7-H4 proteins in different human hepatocellular carcinoma cell lines and to test the inhibitory effect of sorafenib on different human hepatocellular carcinoma cell lines.Results Compared with normal human liver cells (HL-7702),the expressions of B7-H3 and B-H4 proteins in different human hepatocellular carcinoma cell lines were significantly up-regulated (P＜0.01).The cytotoxic activity IC50 values of sorafenib to Hep3B,BEL-7404,MHCC97H,HCCLM3 and HCCLM6 were 14.56,9.14,9.46,17.21 and 9.29 μmol/L respectively.After treating Hep3B,BEL-7404,MHCC97H,HCCLM3 and HCCLM6 with sorafenib at the doses of 5,10 and 20 μmol/L separately,the expressions of B7-H3 and B7-H4 proteins were strikingly down-regulated when compared with the control group (P＜0.01).Conclusion The overexpressions of B7-H3 and B7-H4 proteins in different human hepatocellular carcinoma cell lines are a common finding,which can influence tumor immune escape.It may be a new target for prevention and treatment of liver cancer in future.
ABSTRACT
Objective To investigate the effect of B7-H3 protein,a collaborative signal molecule,on macrophage-inflammatory protein 2 (MIP-2) mRNA level in Streptococcus pneumococcal meningitis mouse models.Methods Forty-eight healthy male BALB/C mice at the age of 1.5-2.0 months were randomly divided into 4 groups:9 g/L saline group(NS group),B7-H3 protein group(B7-H3 group),Streptococcus pneumoniae group(SP group),and Streptococcus pneumoniae plus B7-H3 protein group(combination group),12 mice in each group.Mouse models were established by intracerebral ventricular injection with 9 g/L saline,B7-H3 protein,Streptococcus pneumoniae type 3 or Streptococcus pneumoniae type 3 plus B7-H3 protein.Neurobehavior of different groups was evaluated according to loeffler rule after injection for 6 h and 24 h,then the mice were sacrificed and MIP-2 mRNA levels were tested by Real-time PCR.The results were analyzed by SPSS 18.0 software.Results Neurobehavior scoring results showed that there were no significant differences between B7-H3 group and NS group (P ＞ 0.05) after infection for 6 h and 24 h,while the score of SP group was decreased compared with that of NS group (P ＜ 0.05),and the score of combination group was significantly decreased compared with that of SP group (P ＜ 0.05).Real-time PCR results showed that,compared with the NS group,the relative MIP-2 mRNA level in SP group increased after injection for 6 hours (1.210 ±0.932 vs 1.000 ± 0.008),but the difference was not significant (P ＞ 0.05),while at 24 h post infection,the relative MIP-2mRNA expressions in SP group were significantly increased compared with that of NS group(12.880 ± 7.792 vs 1.000 ±0.091),the difference was significant (P ＜ 0.05).At 6 h post infection,SP + B7-H3 treatment enhanced the MIP-2mRNA production compared to SP infection alone,but the difference was not significant [(1.240 ± 0.804) vs (1.210 ± 0.932)] (P ＞ 0.05) ; while at 24 h post infection,the difference was significant (38.760 ± 6.601 vs 12.880 ± 7.792) (P ＜ 0.05).Conclusion Collaborative signal molecule B7-H3 protein may increase MIP-2 mRNA level in Streptococcus pneumococcal meningitis mouse model,and exaggerate the clinical disease condition.