Researching The Effect Of Lupeol To HepG2Cells

Hepatocellular carcinoma (HCC, primary carcinoma of the liver) is oneof the most common malignant tumors in the world, there are more than500,000new patients each year. The incidence of HCC is very high in china.The disease is characterized by insidious onset has a long incubation period,rapid growth, high malignancy, metastasis and invasion strong, poorprognosis. Currently the most effective treatment method of HCC iscomprehensive treatment, which dominated by surgical resection. However,the majority of patients had lost opportunity to surgery. Only20percentpatients can be treated with surgery. It is reported that the recurrence rate after5years was60-70%. TACE as a secondary treatment was used for the patientswho once undergo surgery or can not tolerate surgery, only6%patients cansurvive after5years. Therefore, a new treatment for liver cancer is eagerlyneeded. Lupoel is a new anti-cancer naturally substances. Recently, somestudies suggest that lupeol can be able to inhibit the growth of various cancers.The effect on liver cancer is rare research report.Objective: Discussion on the influence to HCC proliferation in vivo andin vitro apoptosis of lupeol. Clarifying the effect of anti-hepatoma effect oflupeol.Methods:1Different concentrations of lupeol (20,30,40μmol/L)acts on HepG2cells. The growth inhibition of HepG2cells were detected byMTT assy (24h,48h,72h).2After HepG2cells were incubated different concentrations(0,20,30,40μmol/L), apoptotic and DNA content were detected by Flow Cytomety assy.3We routinely cultured HepG2cells in vitro, and made fore mono-cellsuspension (with1×107/ml cell density and in exponential growth behavior)and subcutaneously implanted on4-week-old nude mices respectively. All themices were divided into4groups. Every group included5mices, and they were subcutaneously implanted with0.2ml mono-cell suspension on theirright nest, it was about2×106cells. Different doses of lupeol (0.25mg,0.5mg,1.0mg) intraperitoneally, Injected once every two days. The state of miaceswere observed. The mices were killed after14days. The volume and weightof the tumors were measured.Results:1MTT assay showed that the different doses of lupeol (20,30,40μmol/L) impact on HepG2cells, after24h cell inhibition rates were2.2%,6.3%,8.6%(P﹤0.05), after48h cell inhibition rates were4.7%,18.6%,23.4%(P﹤0.05), after72h cell inhibition rates were19.5%,23.1%,30.2%(P﹤0.05). Lupeol can inhibit the proliferation of HepG2cells, and the effectwas associated with dose and time.2FCM results showed that the different doses of lupeol (0,20,30,40μmol/L)impact on HepG2cells, after48h apoptosis rates were1.1%,9.7%,15.4%, thedifference was statistically significant (P<0.05). Cell cycle showed that withthe increasing of lupeol, the number of cells which in the S phase increased(increased from27%to45%, P<0.05), in the G2/M phase decreased (from18%down to2%, P<0.05), in the G0/G1phase had no significant(P>0.05).3Vivo test3.1Observing the general of nudesTumor formation rate was100%. Most of the tumors were spherical inthe early, tumors were irregular in shape in the late. The diet, activity andresponse to the outside world of tne nudes had not change significantly in theearly, with the increasing of tumors the nudes showed some unusual behavior,for example eating less, weight loss and unresponsive. There was nosignificant difference (P>0.05) in each group nude body before treatmentquality. After treatment, the control group, low-dose group, middle dose groupand high dose group were body weight (15.41±1.30) g,(16.62±1.46) g,(17.36±0.71) g,(17.84±0.92) g. Compared with the saline group, thehigh-dose group, middle dose group nude body mass increases, the differencewas statistically significant (P<0.05), low-dose group nude body massincrease is not obvious, the difference was not statistically significant. 3.2Volume of tumors in nudesThere was no significant difference in each group tumor volume beforetreatment quality. After treatment, the volume of tumor in the control groupwas (0.52±0.08) cm3, the low-dose group was（0.43±0.05）cm3, themiddle-dose group was（0.37±0.06）cm3, and the high-dose group was（0.33±0.10）cm3, four blocks tumor volume difference was statistically significant(P<0.05).3.3Weight of tumors in nudesAfter treatment, the weight of tumor in the control group was（0.49±0.08）g, the low-dose group was (0.39±0.05)g, the middle-dose group was(0.35±0.06)g, and the high-dose group was（0.29±0.08）g, four blocks tumorvolume difference was statistically significant (P<0.05).Conclusion:1Lupeol can be able to inhibit the growth of HepG2cells,and the effect was associated with dose and time.2Lupeol can be able to induce apoptosis of HepG2cells, and the effectwas associated with dose and time.3lupeol can HepG2cell cycle arrest process.4Intraperitoneal injection of lupeol can inhibit the growth of tumor.5Lupeol will hopefully be used for hepatocellular carcinoma, whichsignificance for exploring a potential treatment strategies.