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The background of 1,25 (OH) 2 VitD 3 normal physiologically active substances in the human body, in recent years, about 1,25 (OH) 2 VitD 3 on the mechanism of action of tumor cells has made some progress, including the inhibition of cell proliferation, reducing tumor cell DNA and RNA synthesis, immune regulation, to reduce tumor angiogenesis and induce apoptosis, but its The exact molecular mechanism has not yet been elucidated. Objective To investigate the 1,25 (OH) to , VitD , 2 3 whether inhibition of retinoblastoma HXO-RB 44 cell proliferation and induced apoptosis reveals 1,25 (OH) 2, VitD , 3 retinoblastoma HXO-RB 44 cells role bax/bcl-2 gene expression. Method 1, grouping: using different concentrations of 1,25 (OH) , VitD , 2 3 treatment of retinoblastoma HXO-RB 44 cells 48 hours, divided into four groups according to the concentration of the drug's different: the high concentration group, 1 × 10 -7 M 1,25 (OH) 2 VitD ; concentration in group 1 × 10 -8 M 1,25 (OH) 2 the VitD 3 ; low concentration group, 1 × 10 -9 M 1,25 (OH) 2 VitD 3 ; the control group, only plus ethanol [1,25 (OH) 2 VitD 3 solvent]. Detection: using the MTT assay spectrophotometer measured OD values; trypan blue staining experiments, live cell rate is calculated by counting board; detect apoptotic cells ratio using flow cytometry (Annexin V-FITC/PI notation); Using semi-quantitative RT-PCR method to detect bax/bcl-2 gene mRNA expression. Results of morphological observation: Compared with the control group of 1,25 (OH) , VitD , 2 3 treatment group retinoblastoma HXO-RB 44 cell volume gradually narrow membrane wrinkles, cell aggregation capacity decreased, increased cell debris; the MTT experiment: 1,25 (OH) 2 the VitD 3 treated retinoblastoma cell tumor the HXO-RB 44 cells OD values ??decreased significantly, the high-concentration group was 0.579 ± 0.084, the concentration in group 0.940 ± 0.098, low concentration group, 1.142 ± 0.097, control group, 1.750 ± 0.162 (p <0.01 vs control); trypan blue experiment: 1,25 (OH) 2 VitD 3 treatment group retinoblastoma HXO-RB 44 cells significantly decreased the number, the high concentration group was 0.83 ± 0.12, in concentrations of 1.28 ± 0.15, low concentrations of 1.53 ± 0.17, control group, 1.96 ± 0.14 (p <0.01 vs control); flow cytometry measurement of apoptosis experiments: 1 25 (OH) 2 VitD 3 treated retinoblastoma cell tumor HXO-RB 44 number of apoptotic cells significantly increased apoptosis percentage of cells to high concentrations of 15.033 ± 1.537% the concentration group 12.866 ± 0.907%, low concentrations of 10.600 ± 1.082%, 2.233 ± 0.839% (p <0.01 vs. control) control group; semi-quantitative RT-PCR was used to detect the bax/bcl-2 gene mRNA expression: 1,25 (OH) 2 the VitD 3 treated retinoblastoma HXO-RB 44 bax gene expression in cells increased, decreased bcl-2 gene expression, of bax/bcl-2 ratio increased, bax gene mRNA relative gray semi-quantitative RT-PCR amplification were high concentrations of 0.699 ± 0.073 in concentration group was 0.539 ± 0.062, low concentrations of 0.472 ± 0.053, control group 0.396 ± 0.034; bcl-2 gene by semi-quantitative RT-PCR amplification of mRNA relative gray were high concentrations of 0.458 ± 0.049, in concentrations of 0.572 ± 0.066, 0.679 ± 0.044 low concentration group, the control group 0.789 ± 0.079; bax/bcl-2 genes by semi-quantitative RT-PCR amplification mRNA expression relative gray were high concentrations of 1.508 ± 0.174, group concentration of 0.944 ± 0.069 low concentrations of 0.695 ± 0.040, control group 0.504 ± 0.053 (p <0.05 vs control). Conclusion of 1,25 (OH) 2 VitD 3 can effectively inhibit the vitro HXO-RB 44 cell growth and proliferation; through streaming cytometry (Annexin V-FITC/PI notation) detection ratio of apoptotic cells confirmed that 1,25 (OH) 2 VitD 3 clear induction HXO-RB < sub> 44 tumor cell apoptosis; preliminary confirmed that 1,25 (OH) 2 VitD 3 to a certain extent raised HXO-RB 44 cells bax gene mRNA expression and down-regulation of bcl-2 gene mRNA expression, possibly through the mechanism of induction of apoptosis in cells HXO-RB 44 .
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