Experimental Study On Apoptosis Inducted By (-)-Gossypo With Combination Of Low Concentration Of Adriamycin In Sof T Tissue Sarcoma Cells And Its Mechanisms

Objective:To investigate Apoptosis Inducted by (-)-Gossypo with combination of low concentration of adriamycin in soft tissue sarcoma cells and Its Mechanisms, offer theoretical basis for the improvement of cancer treatment. Methods:The antiproliferation of drugs on cells was deteceted by MTT assay. The changes of cells morphology and quantity were observed by Wright-Giemsa staining,Hoechst 33258 staining and transmission electron microscopy. DNA ladder formation was determined in Agarose gel electrophoresis.The apoptosis rate and cell cycle of soft tissue sarcoma cells treated by drugs was detected by flow cytometry. And the protein expression of Bcl-2,Bax in soft tissue sarcoma cells treated by drugs were detected by Western-Blot. The changes of caspase-3 and caspase-9 was deteceted by colorimetric assay. Results:(-)-Gossypol and low concentration of adriamycin resulted in a dose and time dependent ihnibition of HT1080 and SW982 cells,(-)-Gossypo with combination of low concentration of adriamycin had more obviouse effet of antiporliefration.(-)-Gossypol and low concentration of adriamycin was found to be bale to induce apoptosis of HT1080 and SW982 cells, as evidenced by typical morphological changes. Agarose gel electrophoresis determined typical DNA ladder formation.Flow cytometry deteceted that (-)-gossypol and low concentration of adriamycin induce apoptosis of HT1080 and SW982 cells (-)-Gossypo with combination of low concentration of adriamycin had more obvious apoptosis-inducing effet. Cell cycle analysis by flow eytometry indicated that (-)-Gossypol and low concentration of adriamycin could change cell cycle.(-)-Gossypol can induce G1 arrest and low concentration of adriamycin can induce S arrest.(-)-Gossypo with combination of low concentration of adriamycin had more obvious increace of S arrest,which evidence that (-)-Gossypol increased the chemotherapy sensitivity of HT1080 and SW982 cells.(-)-Gossypol and low concentration of adriamycin can result in decrease of the protein expression level of Bcl-2 and decrease of the protein expression level of Bax.The combination had more obvious changes. Colorimertie assay showed an activation of caspase-9 and caspase-3. The combination had higher activation changes.Conclusion: In this sutdy,we showed that (-)-Gossypol and low concentration of adriamycin had the effet of antiprolierfation and inducetion of apoptosis on HT1080 and SW982 cells for the first time.(-)-Gossypo with combination of low concentration of adriamycin had more obvious effect. The mechanisms mihgt involve down-relulation of Bcl-2 and up-relulation of bax and affeting cell cycle. Experimental Study on (-)-Gossypo with combination of low concentration of adriamycin in vitro discuss feasibility and interactional mechanism on the effect of (-)-Gossypo with combination of low concentration of adriamycin in soft tissue sarcoma and provide Experimental evidence for screen effective antitumor drugs combining with adriamycin.