Effect of magnesium cantharidate on hepatocellular carcinoma SMMC-7721 cells and subcutaneoushepatocellular SMMC-7721 carcinoma transplantation tumors in nude mice
Author of the article:YAN Rong1, 3** LIU Yun2,3 ZHU Xin-Ting1,3 YI Xiao-Fei1 LIU Liu1 LI Xiao-Fei1, 3***
Author's Workplace：1. Basic Medicine School, Zunyi Medical College, Zunyi 563003, China; 2. Medical and Biological Research Center, Zunyi Medical College, Zunyi 563003, China; 3. Guizhou Provincial College-based Key Lab for Tumor Prevention and Treatment with Distinctive Medicines, Zunyi 563003, China
Key Words：magnesium cantharidate, hepatocellular carcinoma cells SMMC-7721. nude mice, anti-tumor; cell apoptosis
Abstract：[Objectives] To investigate the effect of magnesium cantharidate on hepatocellular carcinoma SMMC-7721 cells and subcutaneous SMMC-7721 transplantation tumors in nude mice. [Methods] 1. The sulforhodamine B(SRB) assay was employed to evaluate the effect of magnesium cantharidate on the inhibition of hepatocellular carcinoma cells in vitro. 2. Flow cytometry（FCM）was used to measure the effect of magnesium cantharidate on cell-cycle arrest and cell apoptosis of hepatocellular carcinoma cells. 3. Hoechst 33342 staining was used to observe the effect of magnesium cantharidate on morphological changes in hepatocellular carcinoma cells. 4. Transmission electron microscopy (TEM) was used to observe the effect of magnesium cantharidate on the ultra-structure of hepatocellular carcinoma cells. 5. A model system using subcutaneous transplantation tumors composed of hepatocellular carcinoma cells was set up in nude mice. The experimental group was injected with magnesium cantharidate (6.26×10-5 mmol) around the tumor and the control group was given the same volume of sterile saline. The inhibition rates of the tumor in the two groups were calculated and compared. 6. Apoptosis in the experimental group was observed with the TUNEL staining method. [Results] 1. The results show that magnesium cantharidate had an obvious inhibitory effect on the proliferation of hepatocellular carcinoma cells, and that this increased gradually with dosage. The IC50 value was 1.79 μmol/L.2.FCM at which point the cell-cycle of SMMC-7721 changed. The ratio of cultured cells declined in the G0/G1 phase, whereas cells increased dramatically in the G2/M phase. The cell-cycle arrested in the G2/M phase. Cell apoptosis increased gradually with concentration of magnesium cantharidate. 3. Hoechst 33342 staining showed that hepatocellular carcinoma cells displayed morphological features typical of apoptotic cells. 4. TEM revealed nuclear abnormities, including concentrations of chromatin on the nuclear edge and the formation of apoptotic bodies. 5. The volume and weight of tumors in the magnesium cantharidate group was significantly lower than in the sterile saline control group (P <0.05) and the inhibition rate was 49%. 6. The TUNEL assay indicated that the apoptosis rate in the magnesium cantharidate group was significantly higher than in the sterile saline group（χ2=92.609, P=0.000）. [Conclusion] Magnesium cantharidate can inhibit the proliferation of SMMC-7721 hepatocellular carcinoma cells both in vitro and in vivo, and induce the apoptosis of cancer cells. Apoptosis may be associated with the arrest of cell division.