Study On Antitumor Activity And Its Mechanism Of Hemsleyin Imine A In Vitro

Uterine cervix cancer is one of malignant tumors of genital system threatening women health and even life,which may be resulted from socioeconomic factor,pathological changes of cervix,virus infection and reduction of immune function,and other facts.The prevention and cure are difficult due to various pathogenesis.At present,the therapy of uterine cervix cancer mostly uses chemosynthesis drugs,such as Cisplatin,Fluorouracil, Ifosfamide,etc.However side effects and multidrug resistance of chemosynthesis drugs is becoming frequent treatment.Natural products have been paid close attention by researcher for their low side-effect and no-drug-resistance.Therefore screening natural products of anti-uterine cervix cancer becomes the study hotspot in pharmacology field.Hemsleya macrocarpa var.clavata is a perennial plant,mainly distributed in southwestern part of China,especially abundant inYunnan and Sichuan provinces.The genus plants have been used as herbal medicines in China for treatment of bronchitis, bacillary dysentery,tuberculosis,gastric ulcer,etc.Furthermore,it also was used for treatment of malignant sore,hypodermal bump by Yunnan minorities.Hemsleyin imine A as a novel imine of sphingolipids was obtained for the first time from Hemsleya macrocarpa vat.clavata.In our study,anti-uterine cervix cancer activity and its action mechanism of Hemsleyin imine A were investigated with Hela cells cultured in vitro.The results were were shown as following.1.Hela cells were treated at different concentrations(320μg/mL,160μg/mL,80μg/mL,40μg/mL,20μg/mL,10μg/mL,5μg/mL)of Hemsleyin imine A at different time points(12hr,24hr,48hr,72hr).They were showed that the differentiation effect induced by Hemsleyin imine A.Under light microscope,the cells appeared cells shrinkage and rounding,cell membrane break,cell float and some cell fragment.These results suggested that Hemsleyin imine A could inhibit growth of Hela cells and induce apoptosis of this cell line.2.The results of MTT demonstrated that with increasing of agent concentration and treatment time,cell numbers were reduced significantly in a dose-and time-dpendent pattern.The 50%inhibiting concentrations(IC₅₀)of 48hr and 72hr were 56.94μg/ml and 23.98μg/ml.3.Annexin V-FITC/PI assay discovered,after being treated with 160μg/mL Hemsleyin imine A for 24hr,48hr and 72hr,that the cells'early stage apoptosis rate were 4.76%,18.6%and 32.1%,respectively,higher than that of controls(p＜0.01);Cell cycle analysis showed the Hemsleyin imine A could reduce the Hela cell proliferation rate with increasing the cell number at G₁/G₀ phase and decreasing the number at S phase and G₂/M phase.All these results suggested that the growth of Hela cells was blocked in G₁→S phases and cell apoptosis was induced by Hemsleyin imine A.All above showed that Hemsleyin imine A has an obvious inhibitory effects against the proliferation and inductive effect to apoptosis to Hela cell in vitro.This study suggested that Hemsleyin imine A might be an anti-uterine cervix cancer and other cancer drug candidate,however,more experiments and investigations in vitro and in vivo are needed.