Inhibition Effect Of 1-methyl-tryptophan On Transplant Hepatocellular Carcinoma Growth In Mice Subcutaneous

Objective:To investigate the Inhibition effect of 1 - methyl-tryptophan (1-MT) on transplant hepatocellular carcinoma growth in mice subcutaneous .Methods:(1) We will construct stably transfected cell lines hepG2 which has IDO gene,and then prove the cell lines by reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot.(2)These mice, which were established human hepatocellular carcinoma subcutaneous tumor models were divided into hepG2 group, empty plasmid group, IDO+ saline group, IDO+ 5 - fluoropyrimidine group , IDO +1-MT group , IDO+ 1-MT + 5 - fluoropyrimidine group ( each group were n = 8).We can observe the mice tumor cases,differences of tumor volume and pathological routine examination, and then detect the expression of IDO in tumor tissues by immunohistochemical .At the same time each mouse was killed before the eyeball blood obtained, separated serum, and then detected the secretion of AFP by ELISA.Results:(1) RT-PCR analysis showed that: pcDNA3.1-IDO group successfully amplified 464bp fragment of the target gene, plasmid pcDNA3.1 group failed to amplify fragments; Western Blot test showed that pcDNA3.1-IDO transfected HepG2 cells express IDO protein, but plasmid pcDNA3.1 group cells and hepG2 cells were no IDO protein.(2)Compared with hepG2 and the empty vector saline group, IDO saline have bulky tumor, fast growth,and the differences were statistically significant (P <0.05); Compared with IDO saline group, 5-FU group ,1-MT group and combined group were smaller tumor volume, less weight, and the tumor inhibitory rates were 86.54%, 79.95%, 94.46% respectively,there were significant differences between them(P <0.05), moreover the combined treatment group seems to be better (P <0.05).But 5-FU group and 1-MT group tumor volume changes were no significant differences (P> 0.05);(3) HE pathological observation: Cell density were decreased and necrotic area were increased of the treatment groups.(4) IDO expression in tumor tissues: Treatment group, cancer cells were less color, smaller and sparse, especially in the combination group there was even no brown color cells; in addition the liver cancer cells were more than the control group, which were arranged in sheets,much larger, colored extensively, and more colored granules.(5) AFP values of peripheral blood in each treatment decreased significantly (P <0.05).Onclusion:(1)Successfully established a stable transfected cell lines hepG2-IDO-gene, and the the IDO gene was expressed effectively in the levels of RNA and protein.We also can further explore the gene function and later laid the foundation for animal experiments.(2) IDO can promote the growth of liver cancer cells which involved in immune escape.(3) 1-MT can inhibite the transplant tumor growth in mice ,and combining 1-MT with chemotherapy ,the therapeuts effect should be better .