The Effect Of ShanXian Granules On Gene Expression Of Bcl-2/Bax In S-180Sarcoma Cells

Objective：This study is to explore the inhibition of the serum containing SD rats SXGin the S-180cells in vitro，the expression of Bcl-2/Bax genes related to thetumor apoptosis, and the mechanism of the anti-tumor occurrence anddevelopment.Method：(1) SD rat serum preparation: randomized the SD rat, administration7days and2hours after the last administration, intraperitoneal injection toanesthesia the rat,then collected the blood though the carotid artery,centrifuge the serum and inactivate the complement,using the microporousmembrane to filter the serum, dispensed and cryopreserved.(2) S-180cells expanded in vitro cell culture are prepared into1×10~5mL~-1cell suspension, which is put into the serum containing SD rat SXG ofdifferent concentrations and the SD rat normal serum. Then, here come thefollowing groups to be divided: Blank group (90%of RPMI-1640medium and10~%FCS), control group (90%of RPMI-1640medium and10~%normal serum of SD rats),serum containing1(10~%of SXG containing serum group and90%RPMI-1640medium),serum containing2(90%of RPMI-1640medium,1%SXG serum containing and9%FCS). MTT assay was used to detect effect of S-180on cell proliferationinhibition rate.(3)According to the group above, each group of three wells, after24hours,48hours,72hours made the cell smears. The expression of Bcl-2/Bax genesin S-180cells is detected through immunocytochemistry (SABC) andimmunofluorescence（FITC），contributing to the comparative analysis. Results:(1)By the each-period comparison between the contorl group and serumcontaing group，the proliferation of S-180cells are significantly inhibit.The proliferation was inhibited obviously，which gradually increased with SXGserum concentation enhanced(P<0.01) and time prolonged(P<0.01).Detect theinhibitory ratio of cell proliferation after72hours in serum containg1is46.46%，which is higer than the serum containg2in the same time.(2) Immunocytochemistry detect S-180sarcoma cells Bcl-2/Bax expression:By the each-period comparison between the control group and blank group,theexpression of S-180cells Bcl-2/Bax genes is not significantly different(P>0.05). Analysis of variance results show that the expression of Bcl-2/Baxgenes is related with time and SXG serum concentration. The Bcl-2expressionis decreased with the rise of the concentrations and prolongation of the time（P<0.01）,while Bax gene increased in S-180cells（P<0.05）.The serum containggroup1and serum containg group2compared have difference（P<0.05）.Thereis difference between the serum group1and group2.(3) By fluorescence microscope after serum effect24hours,48hours and72hours, observed by fluorescence microscopy, the control group and blankgroup showed fluorescent shiny green, granular, and distributed in thecytoplasm and the nuclear membrane. The fluorescence intensity of Bcl-2geneis decreased with the rise of the concentrations and prolongation of the time（P<0.05）. The fluorescence intensity of Bax gene has no significantdifference between the control group and the blank group. It is weekly butvisible（P>0.05）.In the last the fluorescence intensity of serum group issignificant higher than the control group（P<0.05）,and increased with therise of the concentrations and prolongation of the time.Conclusion：The serum containing SD rats SXG significantly causes the expression ofanti-apoptotic gene Bcl-2of the S-180cells in vitro culture to reduce andthe expression of pro-apoptotic gene Bax to increase. Thus the induction of S-180apoptosis attributes to inhibiting its growth.