| Objective: To investigate the efficacy that octreotide combined with cisplatin supressed the growth and proliferation of human ovarian cancer cell line SKOV3 in vitro. and explored the effects of octreotide combined with cisplatin on prolife- ration, apoptosis, morphological alterations and cisplatin sensi- tivity of ovarian carcinoma cell SKOV3 and their possible mechanisms initionally. So as to provide empirical and theore- tical evidence of applying octreotide as the combination with chemotherapy to treat ovarian cancer.Methods:1 Human ovarian cancer cell line SKOV3 was studied in vitro. Different concentration of OCT and DDP were combined in human ovarian cancer cells SKOV3. The growth inhibition ratio of the cells was determined by MTT assay: a. the effects of OCT and DDP alone on proliferation of SKOV3 cells, and the effects of octreotide on proliferation of normal human fibroblast cell line HF; b. the effects of octreotide combined with cisplatin on proliferation of SKOV3 cells; c. the effects of octreotide on the IC50 of cisplatin in SKOV3 cells.2 The reaction of cell proliferation, and the experiment of passage.3 Extracted DNA of SKOV3 cells before and after treated with octreotide were run by agarose gel electrophoresis.4 The distribution changes of cell cycle was analyzed by PI coloration of flow cytometry(FCM) and cell apoptosis ratio was analyzed by flow cytometry.5 The morphological changes of SKOV3 cells were observed by light microscope dyed in Wright-Giemsa.6 The ultromicrostructure of SKOV3 cells were observed by transmission electron microscope.Results:1 The different concentrations of octreotide alone and combined with cisplatin exerted inhibitory effects on proli- feration of SKOV3 cells, and its effect presented a time and dose-dependent way. With the increasing concentration of octreotide and the extending time of treatment, the inhibitory effects were enhanced; Meanwhile, from the results of experiment, we could observe that the inhibition ratio of the combination group was significantly enhanced than the alone group, and there was a significant difference between the treatment group and the control group(P<0.05); The various concentrations of octreotide in combination with cisplatin resulted in the reduction of IC50 in SKOV3 cells.2 The normal SKOV3 cells were in long fusiform or polygon shape, and appearanced fully, grew prosperitly; while the cells treated with octreotide were shrinked and broken, became irregular in shape, the cytoplasm became roughly, floating and cast-off cells increased. The morphological changes became more obviously with the extending of treated time. cispltin had the similar effect with octreotide. The passage exprement sugessted that octreotide combined in HF cells had no reaction, and the SKOV3 cells which treated with octreotide could not passage.3 Extracted DNA of octreotide-treated group cells were analyzed by agarose gel electrophoresis, there were typical ladder-form electrophoresis strips, which means the DNA were degrdned to regular size fragments, while control group cells showed no degrdned fragments.4 After treated with octreotide(1,4ug/ml) for 48h, the number of cells in S phase increased gradually, while the number of cells in G0/G1 phase decreased gradually, which was in dose-dependent way, so octreotide arrested SKOV3 cells in the S phase. and cisplatin arrested SKOV3 cells in the S phase. The effects changed obviously when octreotide combined with cisplatin. Furthermore, after treated with octreotide combined with cisplatin, there were obvious apoptosis ratio of SKOV3 cells, the apoptosis ratio raised up gradually with the increasing concentration of octreotide.5 The morphological changes of SKOV3 cells were observed by light microscope dyed in Wright-Giemsa: SKOV3 cells untreated with octreotide were in long fusiform or polygon shape, and appearanced fully, the nucleus stated in the center of cell, staining well-distributed, and cells grew prosperitly; When treated with octreotide, SKOV3 cells changed significantly in morphology observed by light microscopy, the cells were shrinked and broken, became rounding or orbicular-ovate, the cytoplasm was concentrated, floating and cast-off cells increased. the staining cell nucleus was uneven, there were also caryo-cinesis phenomenon, showing the typical apoptotic morphological changes.6 The ultromicrostructure of SKOV3 cells were observed by transmission electron microscope: the various treatment group of SKOV3 cells showed typical morphology character of apoptosis cell, especially in the combination group. for example, the microvillus became significantly decreased or dispeared, the chromatin concentrated highly, electron density raised and they were enriched as crescent below the nuclear membrane, karyopycnosis could also be seen.Conclusions:1 The different concentrations of octreotide alone and combined with cisplatin exerted inhibitory effects on prolifera- tion of SKOV3 cells, and its effect presented a time and dose-dependent way. So as to provide a theoretical foundation for the clinical use of octreotide to treat ovary cancer.2 The inhibition ratio of the combination group was significantly enhanced than the alone group, and there was a significant difference between the treatment group and the control group (P<0.05); The various concentrations of octreotide in combination with cisplatin resulted in the reduction of IC50 in SKOV3 cells.3 DNA of octreotide-treated group cells were degrdned to regular size fragments, the typical ladder-form electrophoresis strips were shown.4 Octreotide arrested SKOV3 cells in the S phase. and cisplatin also arrested SKOV3 cells in the S phase. The effects changed obviously when octreotide combined with cisplatin. Furthermore, after treated with octreotide combined with cisplatin, there were obvious apoptosis ratio of SKOV3 cells.5 The synergic effects of octreotide combinde with cisplatin of inhibiting proliferation and inducing apoptosis to SKOV3 cells were partly due to change the distribution of cell cycle.
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