Intervention Mechanism Of Modified YangHe Decoction Drug-containing Serum On Breast Caancer MCF-7 Cell Via PI3K/Akt Signal Pathway

Objective:To observe the influence of proliferation,cell cycle,apoptosis and PI3K/Akt signaling pathway related gene and protein expression by Modified YangHe Decoction serum acting on the breast cancer MCF-7 cell,Preliminary discussion on its intervention mechanism of PI3K/Akt signal pathway,lay a foundation of research for the future to explored yang-warming and reduce phlegm method specific mechanism from the angle of cellular level,broaden its application space.Methods:Conventional cell culture and batches of Breast cancer MCF-7 cells,using different concentrations of Modified YangHe Decoction serum to intervene.?1Observed under inverted microscope each drug-containing serum on in vitro cultured breast cancer MCF-7 cell growth condition and form;?2 MTT method to detect proliferation of Modified YangHe Decoction drug-containing serum on MCF-7 cells;?3 FCM detect Modified YangHe Decoction drug-containing serum of MCF-7 cell cycle and apoptosis rate of breast cancer;?4 Scratches and Transwell cell attacks were used to detect the effects of cell invasion metastases;?5 RT-PCR to detect Modified YangHe Decoction drug-containing serum on breast cancer MCF-7 cell PI3K,Akt,caspase-3,CyclinD₁,Bcl-2 and BaxmRNA expression;?6 Western Blot to detect the experession of PI3K,Akt,p-AKT,MMP-2 and TIMP-2 protein.Results:? Inverted microscope visible groups of cells are different degree of morphological change;Modified YangHe Decoction drug-containing serum with high dose group was obviously inhibit the growth of breast cancer MCF-7 cell;? The MTT method shows that,Cell inhibition increased in all groups;? According to the results of FCM,Modified YangHe Decoction drug-containing serum on breast cancer MCF-7 cell after 48 h,middle and high dose group of G₀/G₁stage of cell DNA content decreased significantly?P<0.05?,S phase cell DNA content were significantly increased?P<0.05?;At the same time,early apoptosis rate and late apoptosis rate were higher than the negative control group?P<0.05?;? The results of the attack showed that the breast cancer MCF-7 cells had more membranes and were more aggressive in vitro.After the intervention of the serum containing the serum,the effect of the effect on extracellular invasion was significant.? The results of PCR shows that compared with negative control group,each dose group of cells PI3K,Akt caspase-3,Bax expression values were significantly increased?P<0.05?.On the contrary,the Bcl-2,Cyclin D₁mRNA expression decreased,significant difference between each dose groups?P<0.05?;? According to the results of Western blot,different concentrations of Modified YangHe Decoction drug-containing serum on MCF-7 in breast cancer cells after 24 h,the expression of Akt have no significant change.With the increase of drug concentration,the expression of P13K,P-Akt are decreased,the statistical analysis was carried out on the gray value in the band,compared with the blank group were significantly different?P<0.05?.The high dose group of PI3K,P-Akt content compared with dose group were significantly lower?P<0.05?;At the same time,each dose group of Chinese medicine compound can obviously lower the expression of MMP-2protein?P<0.05?,raising TIMP-2 protein expression?P<0.05?.Conclusion:? Modified YangHe Decoction drug-containing serum can inhibit the growth of breast cancer MCF-7 cell activity,make its different levels of morphological change;? Modified YangHe Decoction drug-containing serum can induce the breast cancer MCF-7 cell had different degrees of apoptosis;? Modified YangHe Decoction drug-containing serum can affect the cell cycle,by blocking cyclinD1 into the nucleus,hence block cell cycle arrest in S phase;? Modified YangHe Decoction drug-containing serum can inhibition of cell invasion and metastasis,be through the regulation of MMP-2/TIMP-2 balance is resisting the invasion and metastasis of breast cancer cells;? The treatment breast cancer resistance mechanism of Modified YangHe Decoction drug-containing serum may be through adjusting the PI3K/Akt signaling pathway and downstream related factors of expression.In order to prevent the proliferation,influence the cell cycle and promote apoptotic and anti-invasion transfer ability.