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| Prokaryotic Expression of Recombinant Protein HS100A6 and Its Biological Effects on Human Osteosarcoma Cell Line 143B |
| ZOU Zheng-yu, WANG Hai-yan, LI Yu-ye, SUN Shuang-shuang, YE Li-wei, WU Rui, DUAN Liang, CHEN Xian, LUO Jin-yong, ZHOU Lan |
| Key Laboratory of Laboratory Medical Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing 400016, China |
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Abstract Objective:To preparation recombinant hS100A6 protein and study its biological effects on human osteosarcoma cell line 143B. Methods:Recombinant plasmid pGST-HRV3C-hS100A6 was constructed and transformed into E.coli BL21, then induced by IPTG. After lysed by ultrasound, the fusion protein GST-HRV3C-hS100A6 was purified by Glutathione-Sepharose 4B beads(GS4BB), next, digested by GST-HRV3C. the pure recombinant hS100A6(rhS100A6) was harvestd after removing GST and GST-HRV3C by GS4BB and then identified by Western blot and quantitatied by Spectrophotometry. MTT assay and Hoechst staining were used to detect proliferation and apoptosis of human osteosarcoma cell line 143B, respectively, transwell was used to detect migration and invasion, Western blot was used to detect expression of β-catenin. Results: rhS100A6 was successfully prepared and the protein yield was about 4 mg/L broth. 30μg/ml of rhS100A6 could promote cell proliferation, migration, invasion and expression of β-catenin(P<0.05), meanwhile had no significant effect on apoptosis of 143B cell line(P>0.05). Conclusion: rhS100A6 is successfully prepared, 30μg/ml of rhS100A6 has promotive effects on human osteosarcoma cell line 143B.
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Received: 28 June 2012
Published: 25 December 2012
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Cite this article:
ZOU Zheng-yu, WANG Hai-yan, LI Yu-ye, SUN Shuang-shuang, YE Li-wei, WU Rui, DUAN Liang, CHEN Xian, LUO Jin-yong, ZHOU Lan. Prokaryotic Expression of Recombinant Protein HS100A6 and Its Biological Effects on Human Osteosarcoma Cell Line 143B. China Biotechnology, 2012, 32(12): 1-7.
URL:
https://manu60.magtech.com.cn/biotech/ OR https://manu60.magtech.com.cn/biotech/Y2012/V32/I12/1
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