The Effect Of Demethoxycurcumin And Bisdemethoxycurcumin To The Invasion And Metastasis Of The Hunman Liver Cancer Cells HepG2

Objective:China has the highest morbidity and mortality in the world. HCC currently ranked No. 2 in the mortality rate of all cancers, and there is a clear upward trend in recent years. Because of the high degree of malignancy, not typical early clinical symptoms, the insidious onset, and rapid progress, metastasis has occurred in most patients, which leads to the loss of the opportunity to line radical surgery, besides, chemotherapy and radiotheropy don't have satisfying efficacy. The occurrence of local invasion and metastasis seriously impact on the prognosis of liver cancer, so an exploratory examination of the mechanism of invasion and metastasis of HCC is of great significance.The process of the tumor invasion and metastasis is a continuous process of the interaction of tumor cells and the host, which concerns several steps, in which the cell adhesion is a very important link, and the cell adhesion molecules plays a important role in the process of embryonic development and differentiation, maintenance of normal tissue structure, inflammation and immune response, wound repair, tumor invasion and metastasis. ICAM-1 and VCAM-1, a member of the Immunoglobulin supergene family, are very important Cell Adhesion Molecules, which exist in the surface of many cells such as vascular endothelial cells, tumor cells, lymphocytes etc. The two molecules are rarely expressed or even not expressed in normal cells, but in tumor cells such as Liver cancer, breast cancer, gastric cancer, renal cancer, colon cancer, they are highly expressed. They can enhance the adhesion between the tumor cells and EMC by combining with their ligands, and then the tumor cells break down the EMC and the basement to promote the tumor invasion and metastasis.Curcuminoids(curcumin demethoxicurcumin and bisdemethoxycurcumin) are a type of pigments isolated from Zingiberaceae turmeric or Curcuma. Researchers have discovered that DMC and BDMC are derivative of curcumin and they have similar pharmacological effects, for example, anti-inflammatory, anti-oxidation, anti-viral, anti-atherosclerosis and anti-tumor. Especially in the anti-tumor aspect, the researchers have found that curcuminoids can dose relatedly inhibit the proliferation of liver cancer, stomach cancer, breast cancer, colon cancer cells, and Induce apoptosis of tumor cells.But curcumin is easily oxidation in vitro, poor solubility in water, is not easy to absorb, bioavailability is not high, limit its wide application.While demethoxicurcumin and bisdemethoxycurcumin is better than curcumin in the stability and bioavailability, there is a good application prospect.Curcuminoids' anti-tumor activity concerns multi-targets, showing that the mechanism is complex, which needs deeper study. As is reported recently, ICAM-1 and VCAM-1 are expressed highly in liver cancer cells and obviously related to the invasion and metastasis of liver cancer, so they can be regarded to be a mark of the invasion and metastasis. Study has confirmed that blocking the function of VCAM-1 and its receptor is benefit to the treatment of ovarian cancer. There are very few reports about whether curcuminoids can inhibit the expression of ICAM-1 and VCAM-1 in the human liver cancer cells Hep G2. The aim of this study is to investigate the possible mechanism of the curcuminoids in inhibiting the invasion and metastasis of the human liver cancer HepG2 cells by researching the effect of the CUR, DMC and BDMC on the expression of ICAM-1 and VCAM-1 of HepG2 cells.Method:1 Hep G2 cells were conventional cultured, different concentrations(10?mol/L?20?mol/L?40?mol/L) of CUR, DMC, BDMC were applied to treat the Hep G2 cells for 24h/48h/72 h, the effect to proliferation of the curcuminoids was detected with MTT assay.2 Various concentrations of CUR, DMC and BDMC were used to interpose the HepG2 cells, and the invasive ability was detected by scratch test and Transwell chamber experiments.3 western-Blot assay was applied to detect the expression of ICAM-1 and VCAM-1 in HepG2 cells after treated by Various concentrations of CUR,DMC and BDMC for 24 hours.Result:1 the results of MTT assay: all of the CUR, DMC and BDMC have the effect of inhibiting the proliferation of Hep G2 cells time-dose- relatedly(P<0.05).2 The scratch test results showed that compared with the 0?mol/L group, the scratch width of the 10?mol/L?20?mol/Land 40?mol/L group were much bigger(P<0.05).3 Transwell chamber experiments results showed that the cells that can passing through the membrane in the 10?mol/L?20?mol/Land 40?mol/L group were fewer than those in the 0?mol/L group(P<0.05). And with the increase of drug concentration,the number of HepG2 cells passing through the membrane significantly reduced, the difference was significant(P<0.05). The three drugs can inhibit the invasion and metastasis of HepG2 cells, and the influence by DMC?BDMC was stronger than CUR.4 Western Blot assay result showed that compared with the control group, the expression of ICAM-1 and VCAM-1 in treated groups were inhibited. The inhibition strength of the ICAM-1 after treated by the same concentration(40?mol/L) of the three kinds of curcuminoids for 24 hours were ranked BDMC>DMC>CUR(P < 0.05), and to the expression of VCAM-1, the influence by BDMC was stronger than DMC and CUR(P<0.05).Conclusion: DMC and BDMC inhibit not only the proliferation but also the invasion and metastasis of the HepG2 cells, and the effect is better than that of curcumin. They can also dose-relatively down regulate the expression of ICAM-1 and VCAM-1 in HepG2 cells(P<0.05).