Experimental Study On Influence Of Bufalin In High Potential Metastatic HCC Cell Lines And The Expression Of Rac1 Protein

BACKGORND & AIMS:Hepatocellular carcinoma (HCC) is one of the most aggressive malignant tumors highly prevalent in China. It transfer easily, there is high recurrence rate after surgical or inerventional therapy. Therefor, to study on the mechanism of invasion and metastasis, and to explore effective treatment of HCC is the challenge in science and clinic. Bufalin is extracted from toxin of Bufo bufo gargarizans Cantor, was reported to have inhibitory effects on HCC. However, the role of anti-metastasis and the underlying mechanism remain elusive. The purpose of this study was to observe influence of bufalin in high potential metastatic HCC cell lines and expression of Racl Protein.METHOD:To study the role of bufalin and relationship in anti-metastasis and the underlying mechanism, we used HCC cell lines (MHCC97H, MHCC97L, LO2) as a based assay system. The inhibitory effect on bufalin of HCC cell's viability and migration was determined by XTT cytotoxicity assay, Scrape migration assay and Cell migration assay. Cell migration and invasion assays were performed by transwell chambers. The expressed of Racl protein in HCC lines, were determined by Western blot.RESULTS:The result of XTT showed that the cell proliferation of MHCC97H was a statistically significant difference compared with MHCC97L/LO2 (P< 0.05). MHCC97L compared with LO2 was showed no significant differece (P< 0.05). The result of Transwell Chamber showed that there was a statistically significant among the three cell lines. The result of Western blot showed that Racl protein did express in the three cell lines (MHCC97H, MHCC97L, LO2). The difference was statistically significant among the three cell lines on expression of Rac1 (P<0.5). Rac1 is highly expressed in aggressive HCC cell lines. Bufalin would reduce the expressed of Racl in aggressive HCC cell lines.CONCLUSIONS:Bufalin could inhibit the proliferation of HCC cell lines (MHCC97H, MHCC97L, LO2), and there showed a time and dose dependent effect. Bufalin could inhibit the migration of HCC lines. And Rac1 was highly expressed in aggressive HCC cell lines. Bufalin could reduce the expressed of Racl in aggressive HCC cell lines. Bufalin may be inhibit MHCC97H cell lines through Rac1 dependent SAPK/JNK pathway.