人Slitrk1基因对PC12细胞增殖和分化的影响

The effect of human Slitrk1 gene on proliferation and differentiation of PC12 cells

目的研究人Slitrk1基因过表达对PC12细胞增殖和分化的影响。方法PCR扩增人Slitrk1CDS,克隆到pcDNA4/myc-HisA载体,构建重组质粒pcDNA4/St1,分别以空载体和重组质粒转染PC12细胞,RT-PCR法鉴定出阳性转染细胞,观察空载体转染的PC12细胞(pcDNA4/PC12)和过表达Slitrk1的PC12细胞(ST1/PC12)的表型并用MTT法检测细胞增殖情况。结果与空载体转染的PC12细胞相比,过表达Slitrk1基因的细胞增殖速度明显减慢。pcDNA4/PC12与野生型PC12细胞表型一致,均为悬浮成团生长,细胞少有突起,而ST1/PC12细胞大部分贴壁生长,突起多见,呈分化状态。结论过表达人Slitrk1基因能够抑制PC12细胞增殖,促进细胞的贴壁及突起长出。人Slitrk1基因可能有促进PC12细胞神经分化的作用。

Objective To study the effect of human Slitrk1 gene on proliferation and differentiation of PC12 cells. Methods The CDS sequence of Slitrk1 was amplified by PCR, and then cloned into pcDNA4 vector. The recombinant plasmid pcDNA4/Slitrk1 were transfected into mt PC12 cells by lipofectamine. The stable expression cell clones were screened by RT-PCR. MTT method was used to detect the proliferation rate of PC12 cells. The morphologic changes in PC12 cells were observed microscopically. Results The stable cell lines expressing pcDNA4 （pcDNA4/PC12） and pcDNA4/Slitrk1 （ST1/PC12） were established. Compared to pcDNA4/PC12 cells, the growth rate of ST1/PC12 cells was decreased. In addition, pcDNA4/PC12 cells tend to grow as well as the normal PC12 cells. However, most of the ST1/PC12 cells adhered to the plate with one or two neurites. Conclusion Over-expression of human Slitrk1 gene inhibited the proliferation of PC12 cells and promoted the outgrowth of neurites. It is suggested that human Slitrk1 gene may be involved in differentiation of PC12 cells.