Study Of7-Hydroxy-5,4’-dimethoxy-2-arylbenzo Furan’s Inhibiting On The Growth Of Cervical Cancer Cells And Its Mechanism

Objective: To investigate cervical cancer cells proliferation areinhibited and apoptosis are induced by drug, and then to investigateits mechanism.Methods:1. MTT assay and soft agar colony formation assay has beenused to detect the proliferation of HeLa cells and Caski cells thatwere inhibited by the drug;2.The Cervical cancer cells wereinoculated under the nude mice’s subcutaneous tissues. To studythe influence of drug on the growth of tumors in nude mice by theestablishment of animal xenograft models;3. Mitochondrial TrackerGreen has been used to detect the integrity of mitochondrialmembrane of the cervical cancer cells and mitochondrial membranepotential has been used to detect the potential changes ofmitochondrial membrane of the cervical cancer cells;4. AnnexinV-FITC/PI double staining has been used to detect the apoptosisrates of the cervical cancer cells after the drug treatment,the DAPInuclear staining had used to observe the morphological changes of the cervical cancer cells after the drug treatment;5. Flowcytometry and Western blot analysis has been used to investigatethe influence of the drug on the distribution of the cell cycle andthe changes of the cells cycle proteins and the related proteins.Results:1.The growth of HeLa and Caski cells treated by the drugwas significantly inhibited compared with the control cells(P<0.05); The experiment of the subcutaneous xenograft tumors in nudemice suggested that the drug could significantly inhibit the growthof the xenograft tumors(P <0.05);2. With the increase of theconcentration of the drug，the morphology of apoptotic cells wasmore obvious and the apoptosis rates of the cervical cancer cellsincreased;3.With the increase of the concentration of the drug，the integrity of mitochondrial membrane of HeLa and Caski cells wasdamaged seriously and the mitochondrial membrane potentialdecreased significantly;4.The drug blocked the cell cycle of thecervical cancer cells in phase S, resulting is increased of CyclinA2,Cyclin E, CDK2and P21,the decreased expression of Cyclin D1,andthe increased protein relation Caspase3, p ATM/ATR and p ERK.Conclusion:1. The drug can significantly inhibit the proliferationof the cervical cancer cells, induce the apoptosis of the cervicalcancer cells by mitochondrial pathway and activate the expressionof Caspase3apoptosis protein;2.The drug can block the cell cyclein S phase and activate the expression of CyclinA2，Cyclin E,CDK2and P21involved in phase S;3.The drug may induce the apoptosisof the cervical cancer cells in phase S by activation p-ERKpathway,so that it can inhibit the growth and proliferation of thecervical cancer cells;4.The drug may repair DNA by activation pATM/ATR pathway in the early stage of the cervical cancer cell DNA damage, but it will induce the apoptosis of the cervical cancercells in phase S,and inhibit the growth and proliferation of thecervical cancer cells.