Screening Chinese Herbal Medicine On Anti-NPC Cells And Study On The Effect Cepharanthine Hydrochiorid On Human NPC Cells And Its Mechanism In Vitro

Background and Objective:Nasopharyngeal carcinoma(NPC),which is threating people's health seriously,is one of the most common malignant tumors in Southern China.The radiation therapy play an important role in early stage of NPC patients,but the five years survival rate is lower.Chemotherapy has the deep significance for late stage of NPC patients,but Chemotherapy often can't be finished because of its serious side effects.Our traditional medicines have the rich experiences in tumors therapy with naturally occurring drugs.The drug action of Chinese herbal medicine,which the side effect is negligible,is mild.However,the synthetic drug has not these features.It is a stringent question to find a kind of Chinese herbal medicine which have more efficient and less side effect on anti-NPC through drug screening.The project group had screened anti-NPC Chinese herbal medicine through two methods.One method is applying blood-serum pharmacology to prepare drug serum and screening them in vitro by MTT.The other method is screening the extractive of Chinese herbal medicines in vitro by MTT.At result,we found scutellariae barbatae serum,Sarcandra serum and the extractive of tea polyphenol,Semen coicis can inhibit the growth of nasopharyngeal carcinoma cells.However,the anti-NPC effect of Sweet Wormwood Herb and Cepharanthine Hydrochiorid(the extractive of Stephania japonica) have been not reported.So our project screen the two Chinese herbal medicines on anti-NPC effect in the same methods as that of the project group,and study on the actions and its mechnism of the Cepharanthine Hydrochiorid(CH) on human nasopharyngeal carcinoma cell lines.Material and Methods:(1) MTT assay had been used to evaluate the growth inhibition of the human nasopharyngeal carcinoma cell line—SUNE-1,CNE-1,CNE-2,5-8F treated with CH and Sweet Wormwood Herb serum,in order to find Chinese herbal medicines whether can inhibit the growth of human nasopharyngeal carcinoma cells;(2) In the human nasopharyngeal carcinoma cell line—SUNE-1,CNE-1,CNE-2,5-8F treated with CH,we used the ATP method to test the vigor,used the flow cytometry to analysis the cell cycle and used the electron microscopes electrophores to observe morphology;(3) DNA microarray had been used to search the different-expressed genes of human nasopharyngeal carcinoma cell CNE-2 before and after treated with CH;And some of different-expressed genes were confirmed by real time-PCR;(4) All experimental data were analyzed with SPSS13.0 for window.RESULTS:(1) MTT results showed that the growth of human nasopharyngeal carcinoma cell line—SUNE-1,CNE-1,CNE-2,5-8F were not obviously inhibited by Sweet Wormwood Herb.The CH could inhibit the growth of human nasopharyngeal carcinoma cell line—SUNE-1,CNE-1,CNE-2,5-8F in vitro conspicuously.There were obviously dosage-effect and time-effect correlation in SUNE-1,CNE-1,5-8F cells,but there was only dosage-effect in CNE-2 cell;(2) ATP method,which results were coherent with the results of MTT, showed CH could inhibit the vigor of four human nasopharyngeal carcinoma cell lines;flow cytometry analysis founded that G₁ arrest and apoptosis peak occurred in the four human nasopharyngeal carcinoma cells treated with CH; Morphological apoptosis of the four human nasopharyngeal carcinoma cells could be observed by means of electron microscope;(3) There were 16450 Unigene tested by DNA microarray in all.There were 138 up-regulated genes and 63 down-regulated genes.According to their biological function,these genes can be classed the genes related to cell proliferation,the genes related to cell apoptosis,DNA repair genes and so on. Parts of genes were confirmed by real time-PCR and the results were corresponded to that of DNA microarray.CONCLUSIONS:(1) The growth of human nasopharyngeal carcinoma cell lines were not inhibited by Sweet Wormwood Herb,but CH could inhibit the proliferation of human nasopharyngeal carcinoma cell lines;(2) CH could arrest the cell cycle of CNE-1,CNE-2,5-8F,SUNE1 at G₁ and inhibite cell proliferation.(3) CH could induce apoptosis of CNE-1,CNE-2,5-8F,SUNE1.impact the growth of the human nasopharyngeal carcinoma cell line-SUNE-1,CNE-1,CNE-2,5-8F obviously and arrest cell cycle in G₀/G₁ and induce apoptosis in vitro;(4)CH can induce the defferent expression of 201genes(138 up-regulated genes and 63 down-regulated genes).(5)The genes which relate to the impact on CNE-2 growth treated with CH can be classed to:the genes relate to cell proliferation,the genes relate to cell cycle,the genes relate to cell apoptosis,DNA repair genes,and so on.we surppose that the molecular mechanism of CH on the inhibition CNE-2 cells growth may be relate to multigenes action.