| Objective To investigate the effect that octreotide combined with oxaliplatinrestrained the growth and proliferation of Human Colon Carcinoma Cell Line SW480invitro and explored the effects of octreotide combined with oxaliplatin on proliferation,apoptosis of Human Colon Carcinoma Cell Line SW480and the possible mechanismsinitially, so as to provide empirical and theoretical evidence of applying octreotide as thecombination with chemotherapy to treat Colon Carcinoma.Methods Human colon carcinoma cell line SW480was studied in vitro. Theexperiment was divided into four groups: control group (no drug group); octreotide group;oxaliplatin group; combination group (octreotide+oxaliplatin). Different concentration ofoctreotide and oxaliplatin alone or combined were used in human colonic carcinoma cellline SW480. The growth inhibition ratio of the cells was determined by MTT assay: theeffects of octreotide and oxaliplatin alone on proliferation of SW480cells, and the effectsof octreotide combined with oxaliplatin on proliferation of SW480cells. DAPI technologywas investigated the apoptosis of colon carcinoma cell; The distribution changes of cellcycle was analyzed by flow cytometry(FCM) and cell apoptosis ratio was analyzed byFCM after using drugs. The gene expression of bax, fas, bcl-2in tumor cells was detecedby semi-quantitative PCR.Results1. The MTT results showed that different concentrations of octreotide oroxaliplatin alone exhibited inhibitory effects on proliferation of SW480cells (P<0.01), andwith the increase of drug concentration, the inhibitory rate also increases gradually. But theinhibitive effect of octreotide did not steadily increase at a dose>25μg/ml. Meanwhile, from the results of experiment, we could observe that the inhibition ratio of thecombination group was significantly higher than oxaliplatin or octreotide alone group, andthere was a significant difference between the treatment group and the control group(P<0.01).2. DAPI assay showed that the nuclear chromatin of the early apoptotic cell waspresentation of condensed state, dyeing deepens, or the nuclear chromatin wascrescent-shaped aggregating at the nuclear membrane side. The nuclear of late apoptoticcells was fragmented into round bodies of varying sizes which was surrounded by the cellmembrane, ie, apoptotic bodies. In the combination experimental group, this phenomenonwas more pronounced (P<0.01).3. Compared with the control group, the cell ratio of DNA synthesis phase (S phase)in the oxaliplatin group was higher, the cell ratio of the G0/G1phase in the octreotide groupwas higher, the difference had statistical significance (P<0.01). Compared with oxaliplatingroup, the cell ratio of DNA synthesis phase (S phase) in the combination group washigher, the difference still had statistical significance (P<0.01). Compared with controlgroup (no drug group) or oxaliplatin group, the effects changed obviously when octreotidecombined with oxaliplatin.4. Apoptosis ratios in the control group, oxaliplatin (12.5μg/ml) group, octreotide(25.0μg/l) group and combination group were6.28±0.23%,10.07±0.53%,10.63±0.14%and15.45±0.41%. Compared with the control group, the apoptosis ratios in theoxaliplatin group, octreotide group and combined group have increased significantly(P<0.05). The apoptosis ratio of the combination group was greater than that of theoxaliplatin group or octreotide group, with statistical significance (P<0.05).5. The semi-quantitative PCR analysis showed that the bax and fas mRNA levels inthe combination group were higher than those in the control group, octreotide group andoxaliplatin group, and the bcl-2mRNA level in the combination group was lower than thatin the control group, octreotide group and oxaliplatin group, the difference had statisticalsignificance (P<0.01). Conclusion Octreotide combined with Oxaliplatin could obviously inhibitproliferation and induce apoptosis of human colon cancer SW480cells in a synergisticmanner, which may be related to the increased bax and fas mRNA, reduced bcl-2mRNAexpression. |
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