The Effects Of Elemene On Survivin And NF-κB Gene Expression And Telomerase Activity In Retinoblastoma HXO-RB₄₄ Cells In Vitro

Retinoblastoma is the most common intraocular malignantneoplasms, it is a big threat to eyesight and lifespan for children.Chemotherapy has become the most important treatment toretinoblastoma in recent years. Side effects and multidrug resistance arethe shortcomings of chemotherapy drugs for retinoblastoma. It isnecessary for us to hunt for new drugs. Elemene (1-methy-1-viny1-2, 4-diisopropenyl-cyclohexane) is a naturally occurring compound that wasisolated from the traditional Chinese medicinal herb, Rhizoma zedoarie.Elemene has direct anticancer effects both in vivo and vitro. Themechanism of its anti-tumor property are not completely understood. Wetreated HXO-RB₄₄ cell with elemene to observe its anticancer activitiesand facilitate the usage of elemene in retinoblastoma treatment.Parâ… The Study on Inhibition of the Growth ofRetinoblastoma HXO-RB₄₄ Cell by Elemene【Objective】To observe the inhibitory effect of elemene onretinoblastoma cell line HXO-RB₄₄ growth【Method】Trypan blue dye exclusion exclusion and MTTcolorimetric assay were used to investigate whether the growth of retinoblastoma cell line HXO-RB₄₄ could be inhibited by elemene or not.Determine the concertration of elemene treating HXO-RB₄₄ cell throughstatistic analysis.【Result】MTT assay show that the growth of HXO-RB₄₄ cellscould be inhibited by elemene significantly and the inhibition depends onthe increase of drag concentration and the elapse of time. The IC50 ofelemene exposed to HXO-RB₄₄ cells for 24, 48 and 72 hours was127.75μg/mL, 85.43μg/mL and 52.95μg/mL.【Conclusion】Elemene can inhibit the proliferation ofretinoblastoma HXO-RB₄₄ cells in vitro, and positively co- related withdose and time. The combination of elemene and VP-16 could promote thesynergistic effect on chemotherapy in the HXO-RB₄₄ cells.【Key words】retinoblastoma, HXO-RB₄₄ cells, elemene, VP-16Partâ…¡Effects of Elemene on both Apoptosis andExpression of Survivin and NF-κB Gene in RetinoblastomaHXO-RB₄₄ Cells【Objective】To investigate the effects of elemene on apoptosis andexpression of Survivin and NF-κB gene in retinoblastoma HXO-RB₄₄cells.【Method】HXO-RB₄₄ cells were divided into four groups after culture, there are a control group and three treatment groups. The latterwere administered by elemene at concentration of 40μg/mL, 80μg/mL and120μg/mL after being incubated for 24h,48h and 72h. Cell cycle andapoptosis were analyzed by flow cytometry (FCM).The expression ofsurvivin and NF-κBp65 were detected with standard immunocytochemistrytechnique and western blot, and the expression of survivin mRNA wasdetected by RT-polymerase chain reaction (RT-PCR). The outputs weresemiquantitative analysised.【Result】HXO-RB₄₄ cells were induced cell cycle arrest byelemene, and the apoptosis cells increased with the increase of dragconcentration and the elapse of time. Elemene could down-regulated thelevel of protein of survivin and NF-κBp65. The effects were be relevant tothe time.【Conclusion】Elemene could induced HXO-RB₄₄ cells apoptosisby down-regulate expression level of survivin and NFκBp65.Partâ…¢The Effect of Elemene on Telomerase Activityand hTERT Expression in Retinoblastoma HXO-RB₄₄ Cells【Objective】To investigate the effect of elemene on telomeraseactivity and hTERT expression, thus to explore the possible mechanism. 【Method】Telomerase activity of HXO-RB₄₄ Cells, treated byelemene, was detected by TRAP assay, and the change of hTERTexpression was observed by western Blotting.【Result】Elemene could down-regulated the expression oftelomerase activity and the level of protein of hTERT. The effects werebe relevant to the time.【Conclusion】Down -regulation of hTERT expression byelemene could inhibite telomerase activity, with a consequence ofinhibition of growth in HXO-RB₄₄ cells. So elemene may be a newchemotherapy drug for retinoblastoma.