Preliminary Study On Anti-tumor Effect Of Sinapine On H22 Mice Hepatocellular Carcinoma And Its Mechanisms

Primary liver cancer(PLC) is one of the most common malignant tumor, which threats seriously to human health and life. Statistics showed that PLC kills an estimated 110 000 people, or forty-five percent of total global deaths each year. Alkaloid extracted from plants is always a very important resources to anti-tumor drugs. However, the drugs that were used extensively in clinic have some defects, such as high toxicity, poor specificity. The development of low toxicity, high efficiency, and good specificity of anti-tumor drugs has become a hot issue in all over the world.In this paper, sinapine was separated from seeds of Brassica alba. The in vitro and in vivo inhibitory effect of sinapine on H22 was studied. And the mechanism of anti-tumor effect of sinapine on H22 hepatoma-bearing mice was explored. These may be the good basis for further study on sinapine and looking for safe and effective anti-tumor drugs.Seeds of Brassica alba were smashed, defatted by ether, extracted by alcohol, and crystallized by potassium thiocyanate, then ethanol and water were used to recrystallize on many occasions. The yield of sinapine was 0.382%. The determination was performed by reversed-phase high performance liquid chromatography (RP-HPLC) and ultraviolet spectrophotometry (UV Spectrophotometry).MTT assay and Trypan blue staining was used to detect the inhibitionrate of H22 hepatocarcinoma cells by sinapine in vitro. And the DNA changes of H22 hepatocarcinoma cells (actived by sinapine) were observed by DNA Ladder. The results showed that sinapine could obviously inhibited the proliferation of H22 hepatocarcinoma cells, and the phenomenon of apoptosis of H22 hepatocarcinoma cells (actived by sinapine) was observed.Subcutaneous injection of H22 hepatocarcinoma cells to mice, creating the mice models of H22. The anti-tumor effect of sinapine was evaluated by observation of rate of tumor inhibition and immunity index. The pathological changes in tumor tissue was observed by HE staining. AO staining, Agarose gel electrophoresis analysis and Immunohisto-chemical assay were used to explore the possible mechanism of induction of apoptosis. The results suggested that the growth of H22 solid tumor in vivo were significantly inhibited by sinapine without toxicity, and the concentration increases with the dose effect relationship. But the effect of sinapine on survivor life of mice bearing H22 ascites tumor is not significant. The immunohisto-chemical results showed that sinapine can enhance the expression of Bax protein and reduced Bcl-2 protein expression.