KIF14在前列腺癌细胞中的表达及作用
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摘要
目的研究驱动蛋白家族成员14(KIF14)在前列腺癌(PCa)细胞中的表达和对PCa细胞的生物学功能的影响。方法采用RT-PCR检测KIF14在PCa细胞系中的RNA表达情况。使用siRNA (siKIF14组或siControl组)分别敲减PCa癌细胞22Rv1,RT-PCR验证敲减效率后,采用细胞计数、Transwell试验分别检测对两侧细胞增殖、迁移的影响。结果 KIF14在多种PCa细胞系中过表达;si KIF14组22Rv1细胞的KIF14的mRNA水平相比siControl组降低,表达量下降达(88±2)%,差异有统计学意义(P<0.05);在KIF14表达被敲减后,siKIF14组细胞增殖相比siControl组在24 h和48 h均降低,差异有统计学意义(P<0.05);Transwell小室试验检测结果显示,24h后,siKIF14组细胞迁移数(139.33±28.74)/每视野,较siControl组的(436.00±51.56)/每视野减少,差异有统计学意义(P<0.05)。结论 KIF14是潜在的癌基因,敲减KIF14表达可抑制PCa细胞的增殖与迁移,可能在PCa进展中发挥作用。
Objective To study the expression of kinesin family member 14(KIF14) in prostate cancer(PCa) cells and its effect on the biological function of PCa cells. Methods The expression of KIF14 in PCa cell line was detected by RT-PCR. PCR cancer cells 22 Rv1 were knocked down by siRNA(siKIF14 group or siControl group), and the knockdown efficiency was verified by RT-PCR. The effects of cell proliferation and migration were detected by cell counting and Transwell assay, respectively. Results KIF14 was overexpressed in various PCa cell lines. The mRNA level of KIF14 in22 Rv1 cells of siKIF14 group was lower than that in siControl group, and the expression level decreased(88 ±2)%, the difference was statistically significant(P<0.05). After KIF14 expression was knocked down, the cell proliferation of siKIF14 group was lower than that of siControl group at 24 h and 48 h,the difference was statistically significant(P <0.05). The results of Transwell chamber test showed that after 24 h, the number of cell migration in the siKIF14 group(139.33 ±28.74)/per visual field was lower than that in the siControl group(436.00 ±51.56)/per visual field,the difference was statistically significant(P <0.05). Conclusion KIF14 is a potential oncogene. Knockdown of KIF14 expression can inhibit the proliferation and migration of PCa cells, which may play a role in the progression of PCa.
Objective To study the expression of kinesin family member 14(KIF14) in prostate cancer(PCa) cells and its effect on the biological function of PCa cells. Methods The expression of KIF14 in PCa cell line was detected by RT-PCR. PCR cancer cells 22 Rv1 were knocked down by siRNA(siKIF14 group or siControl group), and the knockdown efficiency was verified by RT-PCR. The effects of cell proliferation and migration were detected by cell counting and Transwell assay, respectively. Results KIF14 was overexpressed in various PCa cell lines. The mRNA level of KIF14 in22 Rv1 cells of siKIF14 group was lower than that in siControl group, and the expression level decreased(88 ±2)%, the difference was statistically significant(P<0.05). After KIF14 expression was knocked down, the cell proliferation of siKIF14 group was lower than that of siControl group at 24 h and 48 h,the difference was statistically significant(P <0.05). The results of Transwell chamber test showed that after 24 h, the number of cell migration in the siKIF14 group(139.33 ±28.74)/per visual field was lower than that in the siControl group(436.00 ±51.56)/per visual field,the difference was statistically significant(P <0.05). Conclusion KIF14 is a potential oncogene. Knockdown of KIF14 expression can inhibit the proliferation and migration of PCa cells, which may play a role in the progression of PCa.
引文
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[2]Westdorp H,Skold AE,Snijer BA,et al.Immunotherapy for prostate cancer:lessons from responses to tumor-associated antigens[J].Front Immunol,2014(5):191.
[3]Miki H,Okada Y,Hirokawa N.Analysis of the kinesin superfamily:insights into structure and function[J].Trends Cell Biol,2005,15(9):467-476.
[4]She ZY,Yang WX.Molecular mechanisms of kinesin-14 motors in spindle assembly and chromosome segregation[J].J Cell Sci,2017,130(13):2097-2110.
[5]Gruneberg U,Neef R,Li X,et al.KIF14 and citron kinase act together to promote efficient cytokinesis[J].J Cell Biol,2006,172(3):363-372.
[6]Xu H,Choe C,Shin SH,et al.Silencing of KIF14 interferes with cell cycle progression and cytokinesis by blocking the p27(Kip1)ubiquitination pathway in hepatocellular carcinoma[J].Exp Mol Med,2014(46):e97.
[7]Yang T,Zhang XB,Zheng ZM.Suppression of KIF14 expression inhibits hepatocellular carcinoma progression and predicts favorable outcome[J].Cancer Sci,2013,104(5):552-557.
[8]Yang Z,Li C,Yan C,et al.KIF14 promotes tumor progression and metastasis and is an independent predictor of poor prognosis in human gastric cancer[J].Biochim Biophys Acta Mol Basis Dis,2019,1865(1):181-192.
[9]Singel SM,Cornelius C,Zaganjor E,et al.KIF14 promotes AKT phosphorylation and contributes to chemoresistance in triplenegativebreast cancer[J].Neoplasia,2014,16(3):247-256.
[10]Wang Q,Wang L,Li D,et al.Kinesin family member 14 is a candidate prognosticmarker for outcome of glioma patients[J].Cancer Epidemiol,2013,37(1):79-84.
[11]Wang ZZ,Yang J,Jiang BH,et al.KIF14 promotes cell proliferation via activation of Akt and is directly targeted by miR-200c in colorectal cancer[J].Int J Oncol,2018,53(5):1939-1952.
[12]Theriault BL,Pajovic S,Bernardini MQ,et al.Kinesin family member 14:an independent prognostic marker and potential therapeutic target for ovarian cancer[J].Int J Cancer,2012,130(8):1844-1854.
[13]Theriault BL,Corson TW.KIF14:a clinically relevant kinesin and potential target for cancer therapy[M].Kinesins and Cancer,2015:149-170.
[14]Svec D,Tichopad A,Novosadova V,et al.How good is a PCR efficiency estimate:Recommendations for precise and robust qPCR efficiency assessments[J].BiomolDetect Quantif,2015(3):9-16.
[15]Xiao Y,Yuan Y,Zhang Y,et al.CMTM5 is reduced in prostate cancer andinhibits cancer cell growth in vitro and in vivo[J].Clin Transl Oncol,2015,17(6):431-437.
[16]Rhodes DR,Yu J,Shanker K,et al.ONCOMINE:a cancer microarray database and integrateddata-mining platform[J].Neoplasia,2004,6(1):1-6.
[17]Varambally S,Yu J,Laxman B,et al.Integrativegenomic and proteomic analysis of prostate cancer reveals signatures of metastatic progression[J].Cancer Cell,2005,8(5):393-406.
[18]Yu YP,Landsittel D,Jing L,et al.Gene expression alterations in prostate cancer predicting tumor aggression and preceding development of malignancy[J].J Clin Oncol,2004,22(14):2790-2799.