| Objective: To observe the effect of icariin combined with the activity change of calcium sensing receptor(CaSR) on the proliferation of gastric cancer cell SGC-7901 and gene expression of CaSR, Survivin, Runx3, and explore of the mechanism of ICA against tumor, in attempt to provide experimental basis for clinical application of ICA in the treatment of gastric carcinoma. In support of the advantages on effective treatment of gastric cancer in the future and the role in promoting traditional Chinese medicine effective component in antitumor treatment.Methods: Human gastric cancer SGC-7901 cells cultured in vitro1. Effect of ICA and ICA combined with the activity change of CaSR on the proliferation of SGC-7901 cells: Methyl thiazolyl tetrazolium colorimetric assay(MTT) was used to measure the proliferation inhibition rate of SGC-7901 cells which were dispose by increasing dose of ICA(25 mg/L, 50 mg/L, 100 mg/L, 200 mg/L, 400 mg/L) and ICA combined with CaSR agonists and inhibitors after 24 h, 48 h and 72 h.2. Effect of ICA combined with Calindol on apoptosis of SGC-7901 cells: Hoechst staining method was used to test the apoptosis of SGC-7901 cells which were treatment by ICA and ICA combined with Calindol for 48 h.3. Effect of ICA combined with the activity change of CaSR on the protein and mRNA expression of CaSR, Survivin and Runx3 genes in SGC-7901 cells: the cells were treatment by ICA combined with Calindol and Calhex231 for 48 h, and Western blot technology was used to detect protein expression of intracellular CaSR, Survivin and Runx3 genes; RT-PCR technology was used to detect mRNA expression of intracellular CaSR, Survivin and Runx3 genes.4. Statistical methods: All experimental data were analyzed by SPSS17.0. The measurement data followed the normal distribution was expressed by mean value(x±S),variance analysis was used when exceed 3 groups, and SNK pairwise comparison was used if only two groups. If the data does not obey the normal distribution, it was expressed by the median and interquartile, and the rank sum test was used in the comparison between groups.Counting data was expresses by rate, and chi square test was used to compare between groups.All significant level was P<0.05.Results:1. MTT results showed that there had an obvious inhibitory effect on the proliferation of SGC-7901 cells which were treatment by different concentrations ICA for 24 h, 48 h and 72 h.And there was statistical difference compared with the control group(P < 0.05). The inhibitory effect was enhanced with the increase of drug concentrations and prolonged of action time, and showed time and dose dependent manner.2. Regardless of treatment of 24 h, 48 h or 72 h, ICA combined with Calindol inhibition of proliferation of SGC-7901 cells was significantly higher than that only treatment with the same concentration of ICA or Calindol. There was statistical difference compared with the control group(P<0.05), and showed a time dependent manner.3. After Hoechst33342 staining, treatment with ICA combined with Calindol SGC-7901 cells induced apoptosis effect was significantly higher than that only treatment with the same concentration of ICA or Calindol.4. Both ICA and Calindol could promote the expression up-regulation of CaSR, Runx3 protein and mRNA expression, and the expression down-regulation of Survivin protein and mRNA in SGC-7901 cells. After ICA combined with Calindol, the effect of promoting the expression up-regulation of CaSR, Runx3 protein and mRNA, the expression down-regulation of Survivin protein and mRNA was significantly higher than only treatment with any one of the two. And there was statistical difference compared with the control group(P<0.05).Conclusion:1. ICA could obviously inhibit the proliferation of SGC-7901 cells, and showed a time and dose dependent manner.2. The inhibitory effects of treatment with ICA combined with Calindol on SGC-7901 cells were significantly higher than only treatment with any one of them, and showed a time dependent manner.3. ICA could obviously induce the apoptosis of SGC-7901 cells, and the effect of treatment with ICA combined with Calindol on SGC-7901 cells was significantly higher than that only treatment with one of them.4. ICA and Calindol inhibited the proliferation and induced apoptosis of SGC-7901 cellswas related to the expression up-regulation of Runx3 and down-regulation expression of Survivin. |
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