| Objective:CD44 variants include CD44v1, v2, v3, v4, v5, v6, v7, v8, v9, v10, etc., CD44 variant expression in tumor cells in tissue specificity. Currently people think of bone tumors and CD44 expression is closely related to each variant, and that the degree of malignancy of osteosarcoma with CD44 expression variation, invasion, metastasis and so on. Studies have observed tumors of CD44v5, CD44v6 flesh, and CD44v2 relationship with osteosarcoma is not very clear. This study was designed to detect three different osteosarcoma cells by q PCR technology and cell culture(MG-63, U-2OS, Saos-2) expression of the variant CD44v2, CD44v5 and CD44v6, as well as through the cell cycle, growth curve scratch test and judge three different osteosarcoma cell proliferation, migration differences in abilities. Application of statistical methods of correlation analysis to explore the expression of these variants in different osteosarcoma cells, cell growth, proliferation and migration of the relationship.Method: 1. MTT assay, were detected in 1-7 days U-2OS, MG-63, Saos-2 osteosarcoma cell viability, and in turn growth curve; 2 were measured by flow cytometry U-2OS, MG- 63, changes in Saos-2 cell cycle; 3 utilizing scratch test was observed U-2OS, MG-63, Saos-2 cell migration;.. 4 were extracted U-2OS, MG-63, Saos-2 cells of RNA, reverse transcription c DNA, designing q PCR primers were q PCR and fluorescence quantitative detect the expression of three kinds of osteosarcoma cells CD44v2, CD44v5, CD44v6 m RNA; and 5 applications IDEAS software to calculate each gene CT value in the specimens to U-2OS as control standards, applications 2- △△ CT method to calculate the relative quantification of gene expression in the various samples, with mean ± standard deviation(± s) to indicate the relative quantity of each gene in each group of samples, related analysis. Result:1. based on the growth curve showed that MG-63 osteosarcoma cell proliferation, growth rate is higher than the U-2OS and Saos-2, U-2OS osteosarcoma cell proliferation, the growth rate is higher than Saos-2. Osteosarcoma cell MG-63 and Saos-2 compared to P <0.05 was statistically significant.2.CD44v2, CD44v5, CD44v6 have different expression in osteosarcoma cell differences: wherein CD44v2 highest expression in MG-63, U-2OS followed, Saos-2 minimum. CD44v6 expression in osteosarcoma MG-63 cells MG-63 is higher than in U-2OS and Saos-2. CD44v5 and CD44v6 in osteosarcoma cells U-2OS was higher than Saos-2. Inferred osteosarcoma cell migration and cell viability may be proportional to the degree of expression of CD44v2.3. The scratch test results show that cell migration speed from fast to slow as the MG-63> U-2OS> Saos-2.4.q PCR relative quantification showed CD44v2 in osteosarcoma cell MG-63 highest expression, are approximately U-2OS and Saos-2 cells 94 times and 70 times; CD44v5 expression in osteosarcoma cell MG-63, respectively, higher than U-2OS and Saos-2 cells were 7.4 times and 11.1 times; CD44v6 expression in osteosarcoma cell MG-63 were higher than in U-2OS and Saos-2 cells 10-fold and 2.1-fold. CD44v2 also found no significant difference in osteosarcoma cells U-2OS and Saos-2 expression in; the expression of CD44v5 and CD44v6 in osteosarcoma cells U-2OS is higher than Saos-2. Inferred osteosarcoma cell migration and cell viability may be proportional to the degree of expression of CD44v2. Conclusion:1. Three osteosarcoma cells proliferation, migration different: MG-63 proliferation fastest, U-2OS followed, Saos-2 slowest; MG-63 migration fastest, U-2OS followed, Saos- 2 slowest.2.CD44v2, CD44v5, CD44v6 have different expression in osteosarcoma cell differences: wherein CD44v2 highest expression in MG-63, U-2OS followed, Saos-2 minimum. CD44v6 expression in osteosarcoma MG-63 cells MG-63 is higher than in U-2OS and Saos-2. CD44v5 and CD44v6 in osteosarcoma cells U-2OS was higher than Saos-2. Inferred osteosarcoma cell migration and cell viability may be proportional to the degree of expression of CD44v2.3. osteosarcoma cell proliferation, migration may have a different expression of genes CD44v6 CD44v2, CD44v5, related, but the correlation between the needs of these variants further experimental verification. |
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