shRNA誘導(dǎo)的SPERT基因敲低抑制人類結(jié)直腸癌RKO細(xì)胞的增殖并促進(jìn)細(xì)胞凋亡

2018年5月，福建醫(yī)科大學(xué)**附屬醫(yī)院胃腸外科(Department of Gastrointestinal Surgery, The First Affiliated Hospital of Fujian Medical University, Fuzhou, Fujian 350005, P.R. China) LONG-ZHI ZHENG老師研究團(tuán)隊在《ONCOLOGY REPORTS》上發(fā)表論文：

“shRNA?induced knockdown of the SPERT gene inhibits proliferation and promotes apoptosis of human colorectal cancer RKO cells”

“shRNA誘導(dǎo)的SPERT基因敲低抑制人類結(jié)直腸癌RKO細(xì)胞的增殖并促進(jìn)細(xì)胞凋亡”

Abstract：

Colorectal cancer is the third most common type of cancer and the fourth leading cause of cancerrelated deaths worldwide. Although several genes have been identified to contribute to the pathogenesis of colorectal cancer, there are still many genes with unidentified functions in colorectal cancer. This study aimed to investigate the effect of shRNAinduced knockdown of the SPERT gene on the proliferation and apoptosis of human colorectal cancer RKO cells. SPERT was screened based on the TCGA dataset, and SPERT expression, cell growth, proliferation and apoptosis were detected in shSPERT and shCtrltransfected RKO cells. In addition, the SPERTrelated biological pathways were detected using a PathScan? Signaling Antibody Array Kit. We detected lower SPERT expression in shSPERTtransfected RKO cells than in shCtrltransfected cells at both the translational and transcriptional levels (P<0.05), and an MTT assay revealed a clearcut decrease in the proliferation of shSPERTtransfected RKO cells relative to shCtrltransfected RKO cells (P<0.01). A CaspaseGlo? 3/7 assay detected an increase in the caspase3/7 activity and the number of apoptotic cells in the shSPERTtransfected RKO cells than in the shCtrltransfected cells (P<0.01), and flow cytometry detected a higher apoptotic rate in the shSPERTtransfected RKO cells than in the shCtrltransfected cells (20.65±0.26 vs. 5.93±0.06%, respectively, P<0.01). Elevated levels of phosphorylated p44/42 MAPK (ERK1/2), Akt, Bad, HSP27, p38 MARK and Chk2, and elevated PARP and caspase3 expression levels were detected in shSPERTtransfected RKO cells compared with the shCtrltransfected cells (P<0.05). The results of the current study demonstrated that knockdown of SPERT suppresses colorectal cancer cell growth and promotes apoptosis. SPERT may serve as an oncogene and may be a potential target for the treatment of colorectal cancer.

摘要：

結(jié)直腸癌是全球第三大最常見的癌癥類型，也是導(dǎo)致癌癥相關(guān)死亡的第四大原因。雖然已經(jīng)確定了幾個基因參與結(jié)直腸癌的發(fā)病機(jī)制，但仍有許多基因在結(jié)直腸癌中功能不明。本研究旨在探討shRNA誘導(dǎo)的SPERT基因敲低對人結(jié)直腸癌RKO細(xì)胞增殖和凋亡的影響?；赥CGA數(shù)據(jù)集篩選SPERT，在shSPERT和shCtrl轉(zhuǎn)染的RKO細(xì)胞中檢測SPERT的表達(dá)、細(xì)胞生長、增殖和凋亡。此外，使用PathScan?Signaling Antibody Array Kit檢測SPERT相關(guān)的生物學(xué)通路。在翻譯和轉(zhuǎn)錄水平上，科研人員檢測到shSPERT轉(zhuǎn)染的RKO細(xì)胞中SPERT的表達(dá)低于shCtrl轉(zhuǎn)染的RKO細(xì)胞(P<0.05)， MTT實驗顯示，與shCtrl轉(zhuǎn)染的RKO細(xì)胞相比，shSPERT轉(zhuǎn)染的RKO細(xì)胞的增殖明顯減少(P<0.01)。Caspase - Glo?3/7檢測檢測到，與shCtrl -轉(zhuǎn)染的RKO細(xì)胞相比，shSPERT -轉(zhuǎn)染的RKO細(xì)胞的Caspase - 3/7活性和凋亡細(xì)胞數(shù)量增加(P<0.01)，流式細(xì)胞術(shù)檢測到shSPERT -轉(zhuǎn)染的RKO細(xì)胞的凋亡率高于shCtrl -轉(zhuǎn)染的RKO細(xì)胞(分別為20.65±0.26比5.93±0.06%，P<0.01)。與shCtrl轉(zhuǎn)染的RKO細(xì)胞相比，shSPERT轉(zhuǎn)染的RKO細(xì)胞中磷酸化的p44/42 MAPK (ERK1/2)、Akt、Bad、HSP27、p38 MARK和Chk2水平升高，PARP和caspase - 3表達(dá)水平升高(P<0.05)。目前的研究結(jié)果表明，敲低SPERT可抑制結(jié)直腸癌細(xì)胞生長并促進(jìn)細(xì)胞凋亡。SPERT可能是一種致癌基因，可能是治療結(jié)直腸癌的潛在靶點

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