Preliminary Study on Apoptosis of Human Hepatoma Cell Strain MMC7721 Induced by Water-soluble Extract Components of Toad
|School||Dalian Medical University|
|Course||Integrative Medicine Clinical Internal Medicine|
|Keywords||Toad Hepatic carcinoma Apoptosis|
Objective: To observe the toads the water extract (Water-soluble extract components of Toad, WECT) human hepatoma cell line SMMC7721 cell proliferation and apoptosis impact; detection toad aqueous extract of the cell cycle of SMMC7721 cell lines, and The mechanism induced apoptosis preliminary study. Method: the role of water extracts of different concentrations of toad in hepatoma cells SMMC7721, application thiazolyl blue reduction (MTT) detection toad water extract of hepatoma cell line SMMC7721 cells proliferation inhibition and cytotoxicity; application inverted microscope toad the aqueous extract on SMMC7721 cell morphology; Annexin V / PI double staining toad water extract of intervention SMMC7721 apoptosis and necrosis; dUTP nick end labeling, TUNEL assay toad water extract intervention SMMC7721 cells early apoptosis; flow cytometry analysis SMMC7721 cell cycle distribution after drug action situation changes. Results: (1). Toad water extract can inhibit cell division and proliferation, and its half inhibitory concentration of approximately 0.065mg/ml induced apoptosis time, the degree of increase with increasing concentration; (2) toad water extract significant inhibit SMMC7721 cells growth, growth curve showed that the speed of the WECT group of cell proliferation is much slower than in the control group, the the different periods WECT group cell inhibition rate also significantly increased (P <0.05); intervention after the SMMC7721 cells typical pathological changes, cell morphology by polygon into a round, loose chromatin nuclei dissolve, time-dependent morphological changes. (3) apoptosis the WECT process of concentration and time dependent manner, with the increase of the concentration and time extend apoptosis was significantly increased, with the negative control a significant difference (P <0.05). (4) measured by flow cytometry cell cycle change significantly, arrested in S phase of cell division, compared with the negative control group, a significant difference (P <0.05). Conclusion: 1.0.008mg/mL and the above concentration toad water extract human hepatoma SMMC7721 cells with significant cytotoxicity, morphological and DNA fragmentation was confirmed toad water extract-induced cell death to apoptosis; 2 Toad The water extract of HCC SMMC7721 cells proliferation inhibition was concentration and time dependent, 24, 48, and 72-hour half inhibition concentration were: 0.214mg/mL. 0.065mg/mL, 0.023 mg / mL. 3. Toad water extract induced SMMC7721 cell apoptosis, and presents a significant concentration of a time-dependent manner. 4 Toad aqueous extract can inhibit SMMC7721 cell growth, and cell cycle arrest in the S phase, S phase arrest induced liver cancer cell cycle toad aqueous extract may be one of the mechanisms of inhibition of liver cancer cell proliferation.