The Study Of MiR-133b Expression In Liver Cancer Cell HepG2 After Safflower Yellow B(SYB)stimulation

Part I The effect of SYB on HepG2 proliferationObjective: To study the effect of safflower yellow B(safflower yellow B)on the growth and proliferation of liver cancer cells HepG2.Method: Using different concentrations of SYB to act on liver cancer cell line HepG2(Human hepatocellular carcinomas,human liver cancer cells)and cell proliferation has been detected by cell counting kit-8(CCK-8).Different concentration of SYB(0,50,100,150,200 nmol/ml)were added to HepG2 cells respectively,and cultured for 12 h,24h,36 h and 48 h.Final cell number have been counted by CCK-8.Results: HepG2 proliferation has been inhibited by SYB,and inhibition ability also changed with the different concentrations and stimulation time.Statistical results showed that the proliferation ability of HepG2 cells was inhibited significantly when SYB concentration were 0nmol/ml,50nmol/ml,100nmol/ml,150nmol/ml,200nmol/ml,compared with the control group(p<0.05).And when the SYB concentration is150nmol/ml and stimulation time is 36 h,the proliferation ability of HepG2 cells was inhibited most significant.Based on the result above,it is found that the optimal working concentration and time of SYB is 150nmol/ml and 36 h respectively.Conclusion: 1.SYB can inhibit the proliferation of the liver cancer cell line HepG2.2.Different concentrations of SYB have various degree of effects on HepG2 proliferation.3.HepG2 cells proliferation is inhibited significantly when using150nmol/ml SYB to stimulta for 36 h.Thus,find out the optimal SYB concentration for the next experiment.Part II Study the expression of tumor suppressor gene mi R-133 b in HepG2 cells after SYB stimulationObjective: To study the expression changes of tumor suppressor gene mi R-133b(Micro RNA-133 b,micro RNA-133 b fragment)fragment of HepG2 cells after SYB stimulation.Method: Use the optimal inhibitory concentration of SYB(150nmol/ml)to act on the liver cancer cell line HepG2 for different time periods(0,20,40,60 min),and obtain ct DNA(complementary DNA,complementary deoxyribonucleic acid)by using RT-PCR(reverse transcription-polymerase chain reactio,reverse transcription-PCR).The different expressions of mi R-133 b on different time period have been detected by fluorescent quantitative PCR(Polymerase chain reaction,full name polymerase chain reaction).Results: The results found that the expression of mi R-133 b in HepG2 at 20 min,40min and 60 min by SYB were all higher than the expression at 0min,and there was a significant statistical difference(P<0.05).The expression level of mi R-133 b at 40 min and 60 min of SYB was higher than that at 20 min,and there was a significant statistical difference(P<0.05).The expression of mi R-133 b have no significant difference between 40 min and 60min(P>0.05).It shows that the expression level of mi R-133 b has reached the highest level at 40 minutes,and the expression level will not increase when prolonging the time.Conclusion: 1.The expression of tumor suppressor gene mi R-133 b increase when use SYB to stimulate HepG2 cell.2.Through time gradient experiment,it was found that comparing with the control group(0min),the expression of mi R-133 b was significantly increased in experimental group(20,40,60 min),Among them,the expression level of mi R-133 b was the highest at 40 min,and prolonging time could not increase the expression level of mi R-133 b.3.SYB inhibits the growth of HepG2 by increasing the mi R-133 b expression.It is indicated that SYB inhibits tumor cell proliferation by elevating the expression of tumor suppressor gene mi R-133 b,and ultimately inhibits tumor growth.