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[PSF1在结肠癌组织中的表达及其对结肠癌细胞增殖的影响]

[Expression of PSF1 in colon cancer tissues and its effect on the proliferation of colon cancer cells].

作者信息

Wen Ji-zhi, Han Xiao-yan, Wei Bo, Zhang Shi, Wei Hong-bo

机构信息

Sun Yat-sen University, Guangzhou, China.

出版信息

Zhonghua Wei Chang Wai Ke Za Zhi. 2013 Jan;16(1):70-4.

Abstract

OBJECTIVE

To detect the expression of PSF1 (partner of Sld five 1) in colon cancer specimens, and to explore the effect of RNA interference targeting PSF1 on the proliferation of colon cancer cells and its mechanism.

METHODS

Expression level of PSF1 protein in colon cancer specimens was detected by Western blot in 40 patients with colon cancer from May 2004 to December 2006. The short hairpin RNA (shRNA) plasmid targeting PSF1 was transfected into LOVO, HT-29 and HCT116 cells with liposome, then the expression level of PSF1 protein was measured by Western blot, the effect of PSF1 shRNA plasmid transfection on cell proliferation by MTT assay, anchorage-independent growth by soft agar colomy-formation assay, and PSF2, PSF3 and SLD5 mRNA expression by quantitative reverse transcription polymerase chain reaction.

RESULTS

The relative expression level of PSF1 protein in colon cancer tissues was 0.485±0.113, which was significantly higher than that in adjacent normal mucosa tissues (0.056±0.014, P<0.01). Western blot showed that the expression level of PSF1 protein was significantly decreased in colon cancer cells transfected with PSF1 shRNA plasmid. After PSF1 shRNA plasmid transfection, cell proliferation was significantly suppressed, the soft agar colony-forming rates of LOVO, HT-29 and HCT116 cells were significantly lower than those in control groups (P<0.05), meanwhile the expression levels of PSF2, PSF3 and SLD5 mRNA were significantly decreased (P<0.05).

CONCLUSIONS

PSF1 is significantly up-regulated in colon cancer tissues compared with adjacent normal mucosa tissues. ShRNA plasmid targeting PSF1 can inhibit the expression of PSF1 gene, suppress the proliferation of colon cancer cells, suggesting that it may be a new therapeutic target for colon cancer.

摘要

目的

检测结肠癌组织中PSF1(Sld five 1的伙伴)的表达,并探讨靶向PSF1的RNA干扰对结肠癌细胞增殖的影响及其机制。

方法

采用蛋白质免疫印迹法检测2004年5月至2006年12月期间40例结肠癌患者癌组织中PSF1蛋白的表达水平。将靶向PSF1的短发夹RNA(shRNA)质粒用脂质体转染至LOVO、HT-29和HCT116细胞,之后用蛋白质免疫印迹法检测PSF1蛋白表达水平,用MTT法检测PSF1 shRNA质粒转染对细胞增殖的影响,用软琼脂集落形成试验检测其对非锚定依赖性生长的影响,并用定量逆转录聚合酶链反应检测PSF2、PSF3和SLD5 mRNA表达。

结果

结肠癌组织中PSF1蛋白的相对表达水平为0.485±0.113,显著高于癌旁正常黏膜组织(0.056±0.014,P<0.01)。蛋白质免疫印迹法显示,转染PSF1 shRNA质粒的结肠癌细胞中PSF1蛋白表达水平显著降低。转染PSF1 shRNA质粒后,细胞增殖受到显著抑制,LOVO、HT-29和HCT116细胞的软琼脂集落形成率显著低于对照组(P<0.05),同时PSF2、PSF3和SLD5 mRNA表达水平也显著降低(P<0.05)。

结论

与癌旁正常黏膜组织相比,PSF1在结肠癌组织中显著上调。靶向PSF1的shRNA质粒可抑制PSF1基因表达,抑制结肠癌细胞增殖,提示其可能是结肠癌新的治疗靶点。

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