| Objective:To primarily investigate molecular mechanism of diallyl disulfide (DADS) induced CNE2 cell apoptosis, DADS regulated related apoptotic genes was detected by genic microarray in nasopharyngeal carcinoma CNE2 cell.Methods:MTT method detected 50% inhibiting concentration (IC50) that DADS inhibted CNE2 cells proliferation. The cell growth curve analyzed inhibitory action that DADS affected CNE2 cells growth; The apoptosis of DADS affected CNE2 cells checked by Hochest staining, flow cytometry (FCM) and DNA gel electrophoresis. Related genes of DADS induced CNE2 cell apoptosis was detected by genic microarray. The differentially expression of genes and proteins was confirmed by RT-PCR and Western blotting methods.Results:1. IC50 that DADS inhibted CNE2 cells proliferation was 30 mg·L-1. With DADS concentration increased and treatment time extended, CNE2 cells proliferation was obviously inhibited by DADS (P<0.05).2. The result of,Hochest staining showed that 30 mg-L"1 DADS induced typical apoptotic morphology change in CNE2 cells.3. The result of FCM exhibited that apoptosis rate gradually raised as DADS concentration increased (P<0.05).4. The result of DNA gel electrophoresis displayed that there was DNA ladder-shaped when DADS concentration increased to 30mg·L-1.5. Genic microarray together detected 1074 differential expression genes. There was 24 differential expression genes that related cell cycle (19 genes up-regulation expression and 5 genes down-regualtion expression).6. The expression of up-regulated gene SON and down-regualted IL24 detection by genic microarray was verified as same as by RT-PCR and Western blot methods.Conclusion:1. The cellular model that DADS induced CNE2 cell apoptosis was established.2. Apoptosis of DADS induced CNE2 cells was probably related with up-regualtion the expression of TRAF1, CASP9, FOXO1A and IL24 genes and with down-regualtion the expression of PIK3R3, ELMO1, SON and DFFB genes. |
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