Effects Of Jianpi Xiaoji Fang On Migration And Proliferation Of Human Hepatocellular Carcinoma HepG2 Cells And Transplanted Tumor In Nude Mice

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The effects of Jianpi Xiaoji Fang drug plasma on proliferation and migration of liver cancer HepG2 cellsObjective To explore effects on proliferation and migration of liver cancer cells HepG2 influenced by Jianpi Xiaoji Fang drug plasma for further study.Methods The 30 SD rats were randomly divided into two groups,blank control group and treated group with 15 rats in each group.The rats in treated group were administered by gavage 27.2g/kg Jianpi Xiaoji Fang,q12h for 6days,and the rats in control group were treated by normal saline in the same volume.Drug plasma was obtained by filtered and centrifuged.The liver cancer cells HepG2 were cultivated in a period of logarithmic phase and respectively treated with 10%?15%?20%Jianpi Xiaoji Fang drug plasma as treated group And the liver cancer cells HepG2 were treated with rats blank plasma group of the same concentration as blank control group.The HepG2 cells were incubated for 24h,48h and 72h respectively.We used CCK8 assay to explore effects on cell proliferation.And transwell assay was used to explore influences of Jianpi Xiaoji Fang on cell migration.Results CCK8 assay?48h?showed that there was obvious difference between effects on HepG2 cells proliferation of different doses and time of Jianpi Xiaoji Fang drug plasma.(F _group=81.77,P _group=0.000).Optical diversity?OD?values at 24h and 72h were higher than control group of the same concentration?P<0.05?.OD values of 15%drug plasma group and 20%drug plasma group at 48h were lower than control group?P<0.05?.There was a tendency between the same dose groups with time changing in A values(F _time=2996.524,P _time=0.000).And there was interaction effect of groups and time(F _interaction=18.791,P _interaction=0.000).The inhibition rate were merely above 0at 48h.The inhibition rate of 15%drug plasma group and 20%drug plasma group were higher than that of 10%drug plasma group.But there were no significant difference between the three groups?F=1.566,P=0.284?.Transwell assay showed that the migration cells of drug plasma group?97.67±14.20?were lower than that in control group?144.33±10.69?.Conclusions Jianpi Xiaoji Fang drug plasma could be significantly inhibit proliferation and migration of hepatoma HepG2 cells.Part ? The mechanisms on inhibiting liver cancer HepG2 cells growth induced by Jianpi Xiaoji Fang drug plasmaObjective To discuss the biological mechanisms on inhibiting liver cancer HepG2 cells growth induced by Jianpi Xiaoji Fang drug plasmaMethods Cell culture with 15% Jianpi Xiaoji Fang drug plasma was incubated together with liver cancer HepG2 cells as experimental group.Rats blank drug plasma group in the same concentration was used as blank control grou Each group was cultured for 48 hours.RT-PCR was administrated for detection of the gene expression of TGF-?₁ in HepG2 cells influenced by Jianpi Xiaoji Fang drug plasma.The expression of protein of TGF-?₁ gene was detected by western blot.Results After 48 hours,the relative expression of TGF-?₁ mRNA in 15% Jianpi Xiaoji Fang drug plasma group was 0.797±0.023,which is significantly lower than that of control group of 0.994±0.104?t =3.200,P =0.033?.After 48 hours,compared with blank control group,the expression of protein of TGF-?₁ in liver cancer cells HepG2 in 15% Jianpi Xiaoji Fang drug plasma treatment group was significantly lower than that in control group?P < 0.05?.Conclusions The results of RT-PCR and western blot showed that compared with control group,Jianpi Xiaoji Fang drug plasma treatment groups could down-regulate the expression of TGF-?₁ gene and protein of liver cancer cells HepG2?P < 0.05?.Part ? The growth inhibition effects on nude mice induced by Jianpi Xiaoji Fang drug plasmaObjective To establish nude mice transplanted tumor model and explore tumorigenicity of nude mice transplanted of tumor liver cancer HepG2 cells influenced by Jianpi Xiaoji Fang drug plasma in vivoMethods The liver cancer cells HepG2 were cultivated and digested on a period of logarithmic phase.We collected and adapted them with 5×107 cells/ml.A group was treated with 200 ul PBS.B group was treated with 200 ul PBS containing 1880ug/ml Jianpi Xiaoji Fang freeze-dried powder.C group treated with 15% Jianpi Xiaoji Fang drug plasma and PBS,200 ul in total.Those reagents were prepared into HepG2 cell suspension for inoculating nude mice and then they were injected to oxter of nude mice as nude mice transplanted tumor model.The nude mice were feed for 29 days.Then we observed tumor formulation of each nude mouse.They were peeled completely off the skin for tumor organization,weighed and photographed at 30 th days.Tumor formation rate=?the number of tumor formation nude mice-the number of treatment nude mice?×100%.Results Nude mouse transplanted tumor experiment showed that the general condition of nude mice in B and C group was significantly greater than that in A control group.The weight of nude mice in B and C group were 23.34±1.73 g and 22.45±1.23 g,which was significantly lower than that in A group,20.56±2.82g?F =5.19,P =0.015?.But there was no significant difference between B and C group?P =0.91??see Figure 3?.After injection,the weight of tumor in B,C and A groups was respectively 0.91±0.16 g ?0.96±0.17 g ?1.63±0.27 g.The weight of tumor of B and C group were significantly lower than that of control group?F =28.42,P =0.000?.The inhibition rate of transplanted tumor of B and C group were respectively 44.17% and 41.10%.There was no significant difference between B and C group?P =0.63?.Western blot showed that TGF-?₁ was down-regulated in protein level of tumor organism of the nude mice.Conclusions Jianpi Xiaoji Fang drug plasma could suppress tumor formation of liver cancer HepG2 cells in vivo,which accords to the results in vitro.