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Archive > Volume 24 Issue 3 > 2017,24(3):237-241. DOI:10.3872/j.issn.1007-385X.2017.03.004 Prev Next
Basic Research
Effect of Laptm5 3′UTR on proliferation and apoposis of mouse B-cell lymphoma 38B9 line cell
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Abstract:
Objective:To explore effect of lysosomal associated protein transmembrane 5 (Laptm5) 3′untranslated region (3′UTR) on proliferation and apoptosis of mouse B-cell lymphoma 38B9 line cell, by means of establishing B-cell lymphoma cell line with stable overexpression of Laptm5 3′UTR. Methods:Expressions of Laptm5 miRNA and its protein in normal mouse B cell and the mouse B-cell lymphoma 38B9 line cell were respectively examined by Western blotting and fluorescence quantitative PCR. A mouse Laptm5 3′UTR and Laptm5 3′UTR mutation gene fragment containing mutated miRNA 17-3p binding sites were inserted into retrovirus expression vector pMSCV-PIG. The retrovirus was constructed and the mouse B-cell lymphoma 38B9 line cell was infected by the retrovirus. Infection rate of the retrovirus was detected by flow cytometry assay. Cell counting method and flow cytometry assay observed proliferation and apoptosis of the 38B9 cells with Laptm5 3′UTR or mutated overexpression of Laptm5 3′UTR respectively. Results: Expressions of Laptm5 miRNA and its protein in the B-cell lymphoma cell all were more significantly reduced than those in the mouse B cell (all P<0.01). The 38B9 line cell with stable overespression of Laptm5 3′UTR (mutated Laptm5 3′UTR) was succesfully constructed. Proliferation ability of the 38B9 cell with Laptm5 3′UTR was more obviously decreased than that in the 38B9 cell with the mutated Laptm5 3′UTR and apoptosis of the 38B9 cell with Laptm5 3′UTR was more significantly increased than that in the 38B9 cell with the mutated Laptm5 3′UTR （\[7.87±1.08\]% vs \[0.45±0.07\]%, P<0.01). Conclusion: Mouse Laptm5 3′UTR might have functions of inhibiting growth of B-cell lymphoma and promoting its apoptosis. The functions could be associated with regulating its effected-miRNA.
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Project Supported:
Project supported by the National Natural Science Foundation of China (No. 81302041), and the Natural Science Foundation of Jiangsu Province (No. BK20130454)
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