| Objective: Oral squamous cell carcinoma(OSCC)is one of the most common malignant tumors in oral,with a high relapse rate and incidence of lymph node metastasis and poor prognosis,and poses potential serious threats to human health.Ganluqingliangyin is composed of ten kinds of Chinese herbal medicine,including Loquat Leaf,Prepared Radix Rehmanniae,Radix Asparagi,Fructus aurantii,Artemisiae Scopariae,Radix Rehmanniae,Ophiopogon japonicas,Dendrobium,liquorice and Radix Scutellariae,and a variety of its constituents have anti-tumor effect.To study the role of Ganluqingliangyin in OSCC,this experiment selects the DMBA-induced golden hamster buccal carcinoma model to analysis the mechanisms that Ganluqingliangyin inhibit cell proliferation and induce apoptosis of squamous cell carcinoma of buccal mucosa.Methods:Syrian golden hamsters(6-8 weeks old,100 g in weight,male and female)were randomly divided into five groups,10 for the blank control group,without any treatment.The right cheek pouch of the remaining hamsters was injected with 0.3% DMBA in mineral oil two times a week for 12 weeks.Sixty tumor model were then randomly divided into three treatment groups and modle control group(15 in each group).Low dose group,middle dose group and high dose group were fed with 7.5,15,30 g/kg Ganluqingliangyin seven times per week for 4 weeks.At the 17 th week,all the hamsters were sacrificed and cheek pouch tissue was harvested.Flow cytometry was applied to observe the role of Ganluqingliangyin on cell cycles and apoptosis of buccal mucosa squamous cell carcinoma cells.RT-PCR was used to evaluate the expression of Bax and Bcl-2 m RNA in buccal mucosa squamous cell carcinoma cells.All data were used SPSS 13 statistical software,P<0.05 wasstatistically significant.Results:1 After treatment,the percentage of cell cycle in G0/G1 phase of the cells in the low,middle and high dose group,were 45.7±0.3%,49.4±0.5%,62.8±0.6%,respectively;the percentage of cell cycle in S phase were26.1±0.3%,21.5±0.4%,19.7±0.7%,respectively;the percentage of cell cycle in G2/M phase were 28.2±0.4%,29.1±0.3%,17.5±0.4%,respectively.With the increase of drug concentration,the cells increased in G0/G1phase(P<0.01)and decreased in S and G2/M phase(P<0.01)at the same time.2 After treated with Ganluqingliangyin,after testing,flow cytometry,low and high dose group apoptosis rate(hypoploid peak)were 15.02 + 0.26,19.91+ 0.34,30.38 + 0.57,and the modle control group was 11.72 + 0.19,the apoptosis rate(hypoploid peak)gradually increased,the difference is statistical significance.3 After treated with Ganluqingliangyin in low,medium and high dose groups,the expression of Bax m RNA were 1.45±0.38,4.30±0.65,12.52±0.53,respectively;the expression of Bcl-2 m RNA were 3.56±0.22,1.48±0.11,1.02±0.09.The difference was statistically significant(P<0.05),and the expression of Bax and Bcl-2 m RNA was dose-dependent.Conclusions1 Ganluqingliangyin can inhibit cell proliferation,change cell cycle and arrest cell cycle in G0/G1 phase.2 Ganluqingliangyin may be possible to induce apoptosis of buccal squamous cell carcinoma cells by down regulating Bcl-2 expression and up regulation of Bax expression.3 Ganluqingliangyin can inhibit cancer cell proliferation and induce apoptosis of cancer cells,which is expected to become a new drug for the treatment of oral squamous cell carcinoma. |
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