Research The Inhibitory Effect Of Glaucocalyxin B On Human Cervical Cancer Xenografts

BackgroundThe incidence of female reproductive system tumor rising year by year,of which the first is cervical cancer [1].Research shows that human papillomavirus(HPV)infection is a risk factor of cervical cancer,and a survey of the Affiliated Tumor Research Institute of Chinese Science Academy shows that in our country city and countryside HPV virus carrying rate accounted for 15.2% and 14.6% of the population,which means that the prevention of cervical cancer in China should be paid sufficient attention to [2].At present,the treatment of cervical cancer mainly depends on surgery and radiotherapy and chemotherapy,while the surgical trauma,permanent damage to the body of radiation treatment,toxic side effects of chemotherapy drugs limited its scope of application.In recent years,due to the continuous development of traditional Chinese medicine,Chinese medicine treatment with high curative effect and less side effect for the characteristics has become one of the hotspots in the research of tumor area [39];rabdosia japonica belongs to Labiatae,is very common in our country folk,studies have shown that it can inhibite the growth of various tumor cells [40];the active ingredient is mainly terpenoids and flavonoids,Glaucocalyxin A,(GLA)and Glaucocalyxin B,(GLB)is two terpene compound extracted from Rabdosia.Research shows that GLB can inhibit the growth of tumor cells [7],for example: the lung adenocarcinoma AGZY cell,HL-60 cells,human retinoblastoma Rb cells [15-17];previous study showed that GLB can inhibit cervical cancer HeLa and SiHa cell proliferation,but animal studies is not related report.ObjectiveConstruct tumor of cervical carcinoma HeLa and SiHa cells in nude mice,to-explore the effection and mechanism of different concentrations glaucocalyxin B of cervical cancer in animal xenograft,provide a theoretical basis for its clinica-l application.MethodsIn vitro,Hela cells and SiHa cells were transplanted to BALB/C Nu mice in right thigh dorsal subcutaneous,establish nude mice model;when tumor volume is about 0.1cm3.in the tenth day Si Ha group were randomly divided into 4 groups,in the fourteenth day Hela group were divided into 4 groups,respectively.the normal saline control group,10% ethanol group,GLB low dose group and high dose group,5 rats in each group;intraperitoneal injection,HeLa group were given saline 0.4ml/,10% ethanol,GLB0.15umol/g 0.4ml/ only,and 0.20umol/g 0.4ml/only,every two days,a total of 20 days;SiHa group received saline 0.4ml/only and 0.4ml/ of 10% ethanol saline,GLB0.18umol/g 0.4ml and 0.24umol/g 0.4ml/ only,every two days,a total of 20 days.After intraperitoneal injection,at the same time,every 2 days with standard scales weighed and recorded,someone with a vernier caliper accurate measure maximum short and long diameters of tumor,measuring at least two times for average.After injection,before the nude mice were killed,remove the eye to get blood biochemical indexes and tumor markers changes were observed in tumor bearing mice,through tumor growth inhibition test,HE staining,morphology,histological changes were observed in transplanted tumor;through immunohistochemistry and Western-blot assay in transplanted tumor tissue of phosphatase and tensin ho-molog English deleted from chromosome 10,PTEN,Autophagy key regulatory factorBeclin 1,Microtubule associated protein I light English chain 3,LC-3.Results1.The tumor formation rate was 100%.The results showed that in the different concentration of glaucocalyxin B group and control group,the differention of biochem-ical indicators of tumor bearing mice were not statistically significant(P>0.05).Cancer antigen125 and carcinoembryonic antigen in each group of tumor bearing mice with glaucocalyxin B decreased,there was significant difference(P<0.05).The transplanted tumor volume of HeLa and SiHa in the GLB treated group was significantly lower than that in the blank control group(P < 0.05).2.HE staining method showed that GLB treated mice transplanted tumor tissue appeared obvious apoptosis and necrosis change;Immunohistochemistry,Western blotting method showed that GLB treated tumor tissue tumor suppressor gene PTEN,microtubule associated protein LC3II/I,autophagy gene Beclinlprotein expression level is higher than the blank control group(P<0.05).Conclusion1.The growth of HeLa and SiHa cell xenografts in nude mice can be inhibited by glaucocalyxin B,and the concentration dependence.GlaucocalyxinB can be known to lower tumor markers in a concentration dependent manner.Glaucocalyxin B can induce the autophagy of tumor cells,in a concentration dependent manner.2.Glaucocalyxin B could induce tumor suppressor gene PTEN and apoptosis factor Beclin1 and LC3 expression,thus inhibiting cervical cancer cell proliferation and promote apoptosis.