Effects Of Triptlide On Human Oral Cancer Cell Grown And PTEN Expression

Objectives Explore the mechanism of resistance human oral cancer cell to triptolide,through investigating the effect of TP both on the proliferation of human oral cancer cell and expression of phosphates and tension homologue deleted on chromosome ten gene(PTEN),to provide evidence of drug therapy for oral squamous cell carcinoma.Methods As human oral cancer cell a studying object,puting human oral cancer cell as a matched group(A)and four experimental groups(B,C,D,E),respectively give TP dispose on the five groups of 0,1.25,2.5,5.0 and 10μg/L.1 Though MTT staining method to test human oral cancer cell proliferation change affect after 24,48,72 hours with TP;2 Though flow cell measure technique to test human oral cancer cell cycle affect with TP;3 Though Real-time Quantitative polymerase chain reaction to test PTEN m RNA affect after 24,48,72 hours with TP;4 Though Western-blot to test PTEN protein affect after 72 hours with TP.Results 1 The result of MTT: to compare with the matched group,the cell inhibition ratio are(12.48±0.86)%,(23.61±0.10)%,(35.37±0.10)%,(46.58±0.91)%;after A,B,C,D,E groups,difference statistics significance(P<0.05);the cell inhibition ratio are(26.92±0.14)%,(38.67±0.11)%,(72.62±0.89)%,(90.42±0.28)%after A,B,C,D,E groups,difference statistics significance(P<0.05);After 48 h,the experimental group and control group,each group between the two comparison has statistical significance(P<0.05);the result manifests that TP can inhibit remarkably the cell proliferation of human oral cancer cell and present the dependency of concentration and time.2 The result of flow cell measure technique manifest that:after 48 hours of the affect on human oral cancer cell with TP,A,B,C,D,E groups of the G1 cell proportion is(8.78±0.98)%,(70.85 ± 0.41)%,(74.90 ± 0.28)%,(9.14 ± 0.45)%,(84.13 ± 0.47)%;and the S cell proportion is(25.40±0.42)%,(23.14±0.30)%,(20.90±0.25)%,(16.65±0.89),(41±0.73)%;G1 phase and S phase of the control group and experimental group,both two comparison between groups was statistically difference(P < 0.05);it means TP can affect the cycle of human oral cancer cell and there is increase of the dependency of concentration.3 The result of TR-PCR manifest that : after 24 hours,the A,B,C,D,E groups are 1.67,2.22,2.86,3.5 times to the matches groups;after 72 hours,the B,C,D,E groups are 3.95,5.27,7.18,9.16 times to the A groups,Each time point of the control group and experimental group,each group between the two comparison has statistical significance(P < 0.05);it means that TP can up the m RNA express of PTEN,and exist the dependency of concentration and time.4 The result of Western-blot manifest that: after 72 hours on human oral cancer cell with TP,PTEN protein express can up the 3.8,6.8,10.8,17.5times of matched groups,Pairwise comparison between control group and experimental group,each group were statistically significant(P < 0.05);it means that TP can remarkable up the m RNA express of PTEN with the increase of TP concentration,PTEN protein express also manifest regularity increaseConclusions 1 On this experimental conditions,TP can effectively inhibit growth of human oral cancer cell;2 TP can inhibit the human oral cancer cell on the phase G1,thus can decrease the transformation from phase G1 to phase S.3 TP can improve the expression of PTEN’ m RNA and protein in human oral cancer cell.