过表达精子相关抗原9对前列腺癌细胞PC3生物学行为的影响

Effect of over-expression of SPAG9 on proliferation,cell cycle and migration in human prostate cancer cell line PC3

目的研究精子相关抗原9(sperm-associated antigen 9,SPAG9)对前列腺癌细胞增殖、细胞周期、侵袭转移等生物学行为的影响,探讨可能的分子机制。方法前列腺细胞PC3中转染SPAG9后,qRT-PCR和Western blot检测转染后细胞SPAG9的表达变化;CCK-8检测SPAG9对PC3细胞增殖的影响;流式细胞术检测SPAG9对PC3细胞周期的作用;Transwell实验检测转染SPAG9对PC3细胞体外迁移能力的影响。结果 qRT-PCR检测结果表明SPAG9组的SPAG9的相对表达量(20.776 3±3.080 9)明显高于对照组(1.010 2±0.178 4)(P<0.01)。Western blot结果显示在前列腺癌细胞PC3中过表达SPAG9后成功。CCK-8实验表明SPAG9促进细胞增殖;流式细胞术检测结果表明过表达SPAG9使PC3细胞周期加快;在Transwell迁移实验中,SPAG9转染细胞后,SPAG9组穿膜细胞数为(88.078±4.509)/视野,对照组穿膜细胞数为(54.333±4.582)/视野,SPAG9组细胞迁移能力明显高于对照组(P<0.05)。结论 SPAG9促进前列腺癌细胞PC3的增殖、细胞周期及侵袭转移等生物学行为,并可能通过ERK和JNK信号通路发挥这些作用。

Objective To investigate the effect of sperm-associated antigen 9 （SPAG9） over- expression on the proliferation, cell cycle and migration in human prostate cancer cell line PC3 and the relevant molecular mechanism. Methods PC3 prostate cancer cells were transformed with plasmid M29- SPAG9 and plasmid M29, and qRT-PCR and Western blotting were used to detect the SPAG9 expression levels. The cell proliferation was detected by CCK-8 assay. The cell cycle distribution was analyzed by flow cytometry. Transwell migration assay was used to analyze cell migration ability. Results The results of qRT- PCR and Western blot analysis indicated that the SPAG9 expression in the M29-SPAG9 over-expression group was higher than that of the M29 control group （20. 776 3 ± 3. 080 9 vs 1. 010 2 ± 0. 178 4, P 〈 0.01 ）. Compared with the M29 control group, the cell proliferation activity was significantly higher in the SPAG9 over-expression group （ P 〈 0. 05 ） , and migration assay indicated that the cell significantly higher than that of the M29 Conclusion the G2/S phase proportion was increased （P 〈 0. 05 ）. Transwell migration ability of the M29-SPAG9 over-expression group was control group （88. 078 ± 4. 509 vs 54. 333 ± 4. 582, P 〈 0.05 ）. Over-expression of SPAG9 promotes the proliferation, cell cycle and migration in human prostate cancer cell line PC3 through ERK and JNK signaling pathway.