| The human cervical cancer is a malignant disease which threatens women's health severely, whose incidence of cancers in women is second only to breast cancer but mortality is in the first place in gynecological tumors. While the effect of conventional treatment surgery and radiotherapy are not very ideal, and therefore, to explore the pathogenesis of cervical cancer and look for a new chemotherapy treatment of cervical cancer has become a current major public health problem which we are facing with.The essential of tumor's occurence is now generally considered to be cell growth and apoptosis'regulation out of control, and cell growth and apoptosis are mainly controlled through cell cycle. Studies have shown that many tumor suppressor genes such as P53, P21, P27 and others are closely related with the incidence of cervical cancer.Isodon is a kind of perennial herbaceous plants, shrubs or subshrubs, whose main component is diterpenoid. Studies have shown that Isodon diterpenoids have anti-tumor effect, but the specific mechanism has not yet been fully elucidated. Therefore, this study aims to observe the effect of diterpenoid B derived from Rabdosia excise (DB) on human cervical cancer (Hela) cells'growth and its mechanisms,and to provide a theoretical basis and treatment of clues to the therapy of human cervical cancerObjective:To observe the effect of diterpenoid B derived from Rabdosia excise (DB) on the cell line of cervical cancer (Hela) and its related mechanisms.Methods:Hela cell line in culture medium in vitro was treated with different concentrations of DB. MTT assay was used to detect the inhibitory rate of cell growth in different times and different doses,and morphologic changes of cells were observed by phase contrast microscope.The changes of cell cycle are detected by FCM.RT-PCR was exploited to observe the mRNA expressions of P53, P21 and CDK2 mRNAof Hela cells. Immunocytochemical staining was utilized to detect the protein expressions of P53, P21 and CDK2.Results:①MTT results showed that DB can inhibit Hela cell growth in a dose-and time-dependent manner(P<0.05, P<0.01).②We observed the changes of cells under the confocal microscopy, Hela cells after treated with DB showed retarded proliferation and and became round and smaller in size, with some cells floating.③We used FCM to examine cell cycle in experiment,the result revealed that the percentage of Gl phase cells increased obviously after treated with DB,the percentage of G1 phase cells was 64.36%.④After the treatment of DB(2μg/ml,4μg/ml,8μg/ml)for 24h,48h,the mRNA expression of P53 and P21 in drug treated groups are higher than those in control group (P<0.01), meanwhile the level of CDK2 mRNA was less than those in control group (P<0.01).⑤After the treatment of DB(2μg/ml,4μg/ml,8μg/ml)for 24h and 48h,Immunocytochemistry stain showed that the expression level of P53 and P21 protein was up-regulating obviously in drug treated groups,the expression of CDK2 protein was down-regulating obviously (P<0.01)Conclusion:DB can inhibit Hela cell growth in a dose-and time-dependent manner, its mechanism may be related to up-regulating the expressions of P53 and P21, down-regulating the expression of CDK2, and furthermore influencing the transition from G1 to S phase. |
