肿瘤转移抑制基因LASS2/TMSG1全长及其截短体对人前列腺癌细胞生物功能的影响

Effects of full-length tumor metastasis suppressor gene LASS2/TMSG1 and its truncated mutant on the biological function of human prostate cancer cells

目的构建肿瘤转移抑制基因LASS2/TMSG1全长及其截短体质粒,探讨其对人前列腺癌细胞生长、迁移、侵袭等生物学功能的影响。方法构建LASS2/TMSG1基因全长以及3个截短体的慢病毒质粒(pCDH-CMV-MCS-EF1-copGFP-T2A-Puro),并将其稳定转染到人前列腺癌细胞系PC-3M-1E8中,采用Western blot法鉴定稳定转染的效果;然后采用软琼脂集落形成实验、细胞划痕修复试验、Transwell体外侵袭实验等分析LASS2/TMSG1基因全长以及3个截短体对人前列腺癌细胞系PC-3M-1E8生物学功能的影响。结果LASS2/TMSG1蛋白和缺失Homeobox(Hox)结构域的截短蛋白,可以有效抑制PC-3M-1E8细胞的增殖、迁移和侵袭能力(P均<0.001);缺失TLC结构域的LASS2/TMSG1蛋白则丧失抑制前列腺癌细胞生长的功能。结论LASS2/TMSG1蛋白的TLC结构域是抑制前列腺癌细胞生长的关键功能域。

Purpose To construct the full-length and truncated mutant of tumor metastasis suppressor gene LASS2/TMSG1 to explore their effects on cell growth,invasion,migration in human prostate cancer cells.Methods The full-length LASS2 gene and three truncated mutant lentiviral plasmids(pCDH-CMV-MCS-EF1-copGFP-T2A-Puro)were constructed.Human prostate cancer cell line PC-3M-1E8 stably expressed the full length of LASS2 gene and three truncations.Western blot analysis was used to identify the protein expression of stable cell line.We used soft agar colony formation assay,scratch repair experiment,Transwell cell invasion experiment to explore the effects of full length of LASS2 gene and the three truncated mutants on the biological function of human prostate cancer cell line PC-3M-1E8.Results The full length sequence of LASS2 gene and T1 truncated sequence effectively inhibited the proliferation ability,migration and invasion(P all<0.001)of PC-3M-1E8 cells.The LASS2/TMSG1 protein lacking the TLC domain lost its ability to inhibit the growth of prostate cancer cells.Conclusion The TLC domain of the LASS2/TMSG1 protein is a key functional domain that inhibits the growth of prostate cancer cells.