Expression of uc.477+ in mouse B lymphoma cells
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摘要: 目的 根据超保守RNA uc.477+在小鼠正常B细胞与小鼠B淋巴瘤细胞中的表达差异,探讨uc.477+对小鼠B淋巴瘤细胞增殖的影响。 方法 从C57BL/6野生型小鼠脾脏中分选出正常B细胞, RT-PCR检测uc.477+在小鼠正常B细胞与B淋巴瘤细胞中的表达。将uc.477+基因片段构建到pMSCV-PIG逆转录病毒表达载体上,过表达质粒477-pMSCV-PIG和空载体质粒pMSCV-PIG经293T包装成逆转录病毒感染小鼠淋巴瘤细胞38B9, 倒置荧光显微镜和流式细胞术检测感染率。细胞计数和细胞周期检测uc.477+过表达后38B9细胞株和空载体38B9细胞株的增殖变化。 结果 uc.477+在小鼠B淋巴瘤细胞中的表达显著高于小鼠正常B细胞。稳定过表达uc.477+ 38B9细胞株和空载体38B9细胞株构建成功。uc.477+过表达38B9细胞株比空载体38B9细胞株增殖能力强。 结论 超保守RNA uc.477+在小鼠B淋巴瘤细胞中的表达显著高于小鼠正常B细胞中的表达。uc.477+能促进小鼠B淋巴瘤细胞增殖。Abstract: Objective According to the differential expression of ultra-conserved RNA uc.477+ in mouse normal B cells and mouse B lymphoma cells, we designed this study order to explore the effect of uc.477+ on the proliferation of mouse B lymphoma cells. Methods Normal B cells were sorted from the spleens of C57BL/6 mice, and the expression of uc.477+ was examined by RT-PCR in normal B cells and B lymphoma cells of mouse.The uc.477+ gene fragment was inserted into the pMSCV-PIG retroviral expression vector.Overexpression plasmid 477-pMSCV-PIG and empty vector plasmid pMSCV-PIG were packaged into retroviruses via 293T to infect mouse lymphoma cells 38B9, and fluorescence microscopy and flow cytometry were used to examine infection rate.The proliferation change of the 38B9 cell line with uc.477+ over-expression and the 38B9 cell line with empty vector was examined by cell counting and cell cycle. Results The expression of uc.477+ in mouse B lymphoma cells was significantly higher than that in normal mouse B cells.Stable over-expression of uc.477+ 38B9 cell line and empty vector 38B9 cell line were successfully constructed.Proliferation ability of the uc.477+ over-expressing 38B9 cell line was more stronger than the empty vector 38B9 cell line. Conclusion The expression of uc.477+ in B lymphoma cells was significantly higher than that in normal B cells of mouse.The uc.477+ can promote the proliferation of B lymphoma cells.
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Keywords:
- ultra-conserved RNA /
- B lymphoma cell /
- 38B9 cell line /
- cell proliferation /
- non-coding RNA
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