| Objective Matrine is one of the active ingredients—an alkaloid from the extract of traditional Chinese medical herb sophora flavesscens Ait. root. Matrine had been reported to have various pharmacological functions. Our long-term research had proved that matrine could significantly inhibit the proliferation of human leukemic cell lines K562 and induce K562 cells differentiate into normal morphological cell in vitro. Recently, there are increasing study on the matrine's inhibitory effects on tumor cell lines other than K562 cells, and more attention has been paid to its anti-tumor effects both in vitro and vivo. Nowadays, exploiting our Chinese medicine treasury, excavating common traditional medical herbs and endowing them up-to-date pharmcological explication has been thought to be the general trend of Chinese medicine development. Now, many people are devoting to develop new anti-cancer pharmaceuticals from Chinese herbs medicine, which is necessary to carry forward our precious traditional Chinese medicine. Under this background, it is undoubtedly a promising and singnificant work to investigate thorough the inhibitory effects of matrine on tumor, especial on solid-tumor, and explain its mechanism of anti-tumor effects in vitro and in vivo. Ginven that, we investigated the inhibitory effects of matrine in vitro and in vivo with cell cultivating method, H22 xenograft tumor models estabilishing in BALB/C, and explored the probable mechanisms of matrine in tumor inhibition. We also observed the effects of matrine on the BALB/C's immunofunction in vivo, trying to analyze the mechanisms of anti-tumor effects of matrine and bring them to the light from immunological perspective. Owing to the immunological response lower in tumor-bearing body, we constructed the eukaryotic expression vector pIRES-EGFP-TIM2 with a murine co-stimulation molecule gene—TIM2 gene inserted and transfected it into H22 hepatocarcinoma cells by Lipofectamin 2000. After electing by G418 pressure and limited dilution method in turn, we obtained the monoclone of the positive H22 cells stably espressing TIM2 gene mRNA and EGFP protein in vitro named as H22-TIM2. We regarded it as the murine H22 hepatocarcinoma whole cell vaccination applied to perform the following experiment in BALB/C. The H22-TIM2 cells were subcutaneously inoculated into BALB/C, expecting improving the immunogenecity of H22 cells, and established the tumor-bearing mice models again. Then we researched the inhibitory efficacy of matrine under the immunofunction improved in this model mice. Our data may provide important clues to clinic application of matrine in tumor treatment in the future. Methods 1. Taking the murine hepatocarcinoma cell line H22 as our experimental object, we observed the H22 cell's proliferation inhibition treated with matrine and discussed the probable mechanisms of anti-tumor effects of matrine in vitro from the following aspects: the morphological changes of H22 cells under the optical microscope was observed; changes of cell number,the inhibitory rate of cell proliferation and cell growth curve were detected using MTT assay and cell counting method, respectively; cell necrosis was manifested with trypan blue staining method;the effection of matrine on the cell cycle of H22 cells was examined with fluorescein activated cell sortor (FACS) and the cell apoptosis induced by matrine at the early stage was detected with Annexin Ⅴ-FITC/PI double marked assay; the expression of Bcl-2 and Bax protein in H22 cells were detected using immunohistochemistry assay. 2. H22 xenograft tumor models were established in BALB/C. The BALB/C mice in experimental groups were administed matrine with 50mg/kg weight and 100mg/kg weight dosage through abdominal cavity injection, respectively; while those in the control groups were administed 20mg/kg weight CTX and N.S solution, respectively. Matrine were injected into BALB/C mice 7 times q.o.d. The tumor suppression effects of matrine was studied in vivo from the following aspects: the xenograft tumor were excis...
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