Sinomenine Inhibit Proliferation Of MGC803Gastric Adenocarcinoma Cells Via Arresting Cell Cycle And Affecting The Apoptosis

【Objective】Study the inhibitory effects of MGC803gastric adenocarcinoma cellsproliferation by sinomenine and dicuss its mechanism【Methods】1. MGC803gastric cancer cells which were cultured in vitro were collected anddivied into five groups，including control、SIN(300、600、900、1200ug/ml) groups.The cells of five groups were distinguish treated with DMEM、SIN(300、600、900、1200ug/ml)for24、48、72hours. Ahft that, The cells were analyzed by CCK8assay forits relative viability. the inhibitory rates of cells were calculated by formula.2. MGC803gastric cancer cells which were cultured in vitro were collected anddivied into five groups，including control、SIN(300、600、900、1200ug/ml) groups.The cells of five groups were distinguish treated with DMEM、SIN(300、600、900、1200ug/ml)for48hours. After that, the cells cycle distribution was determined byflow cytometry (FCM). The expression of p21was determined by Western Blotassay.3. MGC803gastric cancer cells which were cultured in vitro were collected anddivied into five groups，including control、SIN(300、600、900、1200ug/ml) groups.The cells of five groups were distinguish treated with DMEM、SIN(300、600、900、1200ug/ml)for48hours. After that, quantitative analysis of apoptosis by flowcytometry (FCM) was assessed using an Annexin V FITC/PI Apoptosis Assay. Theexpressions of Cyt C in the cystol and mitochondria、Bax、Bcl-2、Clevead Caspase-3、Survivn were determined by Western Blot assay. 【Results】1. Sinomenin inhibited the proliferation of MGC803gastric cancercell.Compared to the blank control group, the inhibition ratios of SIN groups (600、900、1200ug/ml) of24hours（7.71±1.70、15.42±3.95、35.75±2.61）%increasedsignificantly(P<0.05); SIN group(300ug/ml)（2.25±1.14） showed no significantdifference; Compared to the blank control group, the inhibition ratios of SIN groupsof48hours(10.36±2.82、18.02±1.01、24.36±2.65、40.59±3.18)%increasedsignificantly (P<0.05); Compared to the blank control group, the inhibittion ratios ofSIN groups of72hours(19.57±1.36、26.31±3.34、44.00±3.96、53.25±5.51)%increasedsignificantly(P<0.05).2.1Sinomenine affected the cells cycle distribution.Compared to the blank control group（61.13±1.22）%, the G1phases of cells ofSIN groups （70.39±0.31、69.22±0.36、83.35±4.02、87.15±2.82） increasedsignificantly(P<0.05); Compared to the blank control group（26.46±1.02）%, the Sphases of SIN groups（20.03±2.22、17.65±1.82、9.87±1.26、9.33±0.82）%decreased.Compared to the blank control group（12.41±1.45）%, the G2/M phases of SIN groups（9.57±0.68、8.03±1.06、6.78±1.07、3.52±0.92）%decreased.2.2Sinomenine affected the expression of p21.Compared to the blank control group(1573±61.10) INT×mm, the expression ofp21of SIN groups(1806±111.70、2610±130.00、2923±150.44、3717±155.03)INT×mmincreased significantly(P<0.05).3.1Sinomenine induced apoptosis.Compared to the blank control group（0.12±0.03）%, the apoptosis rates of SINgroups （2.49±0.51、4.25±0.67、10.35±1.61、12.82±1.41）%increasedsignificantly(P<0.05). 3.2Sinomenine affected the expression of Cytc in the cystol andmitochondria、Bax、Bcl-2、Clevead Caspase-3、Survivn.Compared to the blank control group(1573±61.10) INT×mm, the expression ofCytc in the cystol of SIN groups (1806±111.70、2610±130.00、2923±150.44、3717±155.03)INT×mm increased significantly(P<0.05); Compared to the blankcontrol group((1623±135.77) INT×mm, the expression of Cytc in the mitochondria ofSIN groups (981±33.49、831±32.70、644±78.14、660±98.49)INT×mm decreasedsignificantly(P<0.05).Compared to the blank control group(794±53.78) INT×mm, the expression of Baxof SIN groups(1314±60.89、1757±70.24、2286±135.0、2557±84.54)INT×mmincreased significantly(P<0.05); Compared to the blank control group(2153±70.50)INT×mm, the expression of Bcl-2of SIN groups (600、900、1200ug/ml)(1640±105.35、1009±29.57、714±73.50)INT×mm decreased significantly(P<0.05), SIN group(300μg/ml)(2150±105.36) showed no significant difference.Compared to the blank control group(1621±38.50) INT×mm, the expression ofClevead Caspase-3of SIN groups(2075±155.93、2523±116.76、3605±168.84、4015±132.57)INT×mm increased significantly (P<0.05).Compared to the blank control group（2990±160.93）INT×mm, the expression ofSurvivin of SIN groups（2313±123.42、1857±150.11、1167±150.44、582±151.19）INT×mm decreased significantly (P<0.05).【Conclusion】1. Treatment with Sinomenine significantly inhibited the growth of MGC803gastriccancer cells in a concentration-and time-dependent manner.2. Treatment with Sinomenine induced the G1phase arrest of MGC803gastric cancercells involved upregulating the expression of p21.3. Treatment with Sinomenine induced apoptosis of MGC803gastric cancer cellsinvolved upregulating the expression of Cytc in the cystol、Clevead Caspase-3andthe value of Bax/Bcl-2and downregulating the expression of Cytc in the mitochondria、Survivn.