ACSL3在前列腺癌细胞系中的表达及其对前列腺癌转移的影响

 目的:观察比较长链脂酰辅酶A合成酶3（long-chain acyl-CoA synthetase 3, ACSL3）在前列腺正常上皮细胞和不同类型前列腺癌细胞系中的表达差异，通过构建ACSL3基因稳定表达的前列腺癌细胞株研究ACSL3对前列腺癌细胞侵袭能力的影响。方法:利用RT-PCR方法检测ACSL3 mRNA在前列腺正常上皮细胞、局限性及转移性前列腺癌细胞中的表达情况，分析其表达与前列腺癌发生、进展及转移的关系；以前列腺癌细胞基因组cDNA为模板，通过PCR扩增ACSL3基因，重组构建含有Flag标签的pCDNA3.1(+)-Flag-ACSL3质粒，包装慢病毒，转染前列腺癌细胞，通过荧光显微镜、RT-PCR和Western blotting等方法鉴定ACSL3在细胞内的表达情况；利用Transwell侵袭实验研究ACSL3对前列腺癌细胞侵袭能力的改变。结果:较前列腺正常上皮细胞，各种前列腺癌细胞中ACSL3 mRNA表达均明显下降。同时，局限性前列腺癌细胞中ACLS3 mRNA表达较转移性前列腺癌细胞明显升高；此外，雄激素非依赖性前列腺癌细胞比雄激素依赖性前列腺癌细胞中ACLS3 mRNA表达显著降低。进而，我们成功构建和包装了表达ACSL3基因的pCDNA3.1(+)-Flag-ACSL3重组质粒及慢病毒，并转染前列腺癌细胞，筛选出稳定表达株；并且，通过Transwell实验证实ACSL3表达与前列腺癌细胞的侵袭能力呈负相关。结论:前列腺正常上皮细胞及不同前列腺癌细胞系中ACSL3的表达存在明显差异，提示ACSL3可能在前列腺癌的发生发展中起重要作用；成功构建了ACSL3基因稳定表达的前列腺癌细胞株，并初步证实ACSL3过表达可以抑制前列腺癌细胞的侵袭力。

Role of ACSL3 expression in suppressing prostate cancer cell metastasis

AIM:To analyze the difference of long-chain acyl-CoA synthetase 3 (ACSL3) expression between normal prostate epithelial cells and prostate cancer cells.METHODS:ACLS3 mRNA expression between normal prostate epithelial cells and prostate cancer cells was compared using RT-PCR. Meanwhile, ACSL3 gene was amplified from prostate cancer cells, and the eukaryotic expression plasmid pCDNA3.1(+)-Flag-ACLS3 and lentivirus Lenti-ACLS3 were constructed. After transfection of ACSL3-plasmid and lentivirus into the prostate cancer cells, ACSL3 expressive was detected by RT-PCR and Western blotting, and then Matrigel invasion assay was performed to investigate the alteration of the invasive ability of the prostate cancer cells with over-expression of ACSL3. RESULTS:A significant difference of ACSL3 mRNA level between normal prostate epithelial cells and prostate cancer cells was observed. ACSL3 was highly expressed in localized prostate cancer cells compared to metastatic prostate cancer cells, while ACSL3 expression was higher in androgen-dependent prostate cancer cells than that in androgen-independent prostate cancer cells. Furthermore, the eukaryotic expression plasmid and lentivirus containing ACLS3 gene were successfully constructed. The prostate cancer cell line which stably over-expressed ACLS3 was established. Up-regulation of ACSL3 inhibited the invasive ability of prostate cancer cells. CONCLUSION:ACSL3 plays an antagonistic role in invasiveness of prostate cancer.