S1 RNA結(jié)合域1的沉默抑制人非小細(xì)胞肺癌細(xì)胞生長(zhǎng)并促進(jìn)細(xì)胞凋亡

2019年1月，北京協(xié)和醫(yī)學(xué)院國(guó)家腫瘤中心/國(guó)家腫瘤臨床研究中心/中國(guó)醫(yī)學(xué)科學(xué)院腫瘤醫(yī)院放射腫瘤科；北京環(huán)興腫瘤醫(yī)院放射腫瘤科(Department of Radiation Oncology, National Cancer Center/National Clinical Research Center for Cancer/Cancer Hospital, Chinese Academy of Medical Science, Peking Union Medical College, Beijing 10021, China；Department of Radiation Oncology, Cancer Hospital of Huan Xing, Beijing10021, China) Tao Zhang老師研究團(tuán)隊(duì)在《Translational lung cancer research》上發(fā)表論文：

“Silence of S1 RNA binding domain 1 represses cell growth and promotes apoptosis in human non-small cell lung cancer cells”

“S1 RNA結(jié)合域1的沉默抑制人非小細(xì)胞肺癌細(xì)胞生長(zhǎng)并促進(jìn)細(xì)胞凋亡”

Abstract：

Background: To investigate the expression of S1 RNA binding domain 1 (SRBD1) in non-small cell lung cancer tissue and the effects of SRBD1 silencing on the biological behaviors of human non-small cell lung cancer cells, and to explore the molecular mechanism of SRBD1functions in human non-small cell lung cancer cells.

Methods: Expressions of SRBD1 in human non-small cell lung cancer tissues and cell lines were examined by immunostaining and RT-PCR. shRNAs of SRBD1 were chemically synthesized and transfected into A549 and NCI-H1299 cells by lentivirus. Cell proliferation was assayed by cell counting, MTT and clone formation. Cell apoptosis was assayed by flow cytometry. Tumorigenicity was assessed by cell injection into BALB/c athymic nude mouse. Gene chip analysis was employed to explore genomic changes in A549 cells. Potential classical signaling pathways, upstream regulators and gene interaction networks were analyzed by Ingenuity Pathway Analysis, and verified by western blot analysis.

Results: SRBD1 was specifically expressed in human squamous cell carcinoma and highly expressed in lung cancer cell lines, NCI-H1299, A549 and NCI-H1975. SRBD1 directed-shRNA (shSRBD1) effectively reduced the expression of SRBD1 in A549 and NCI-H1299 cells. SRBD1 silencing inhibited cell proliferation, and promoted cell apoptosis in non-small cell lung cancer cells, and suppressed tumorigenesis in a nude mouse model. In addition, we found silencing of SRBD1 expression resulted in marked changes in gene expression in A549 cells. Besides, in shSRBD1 group, the protein levels of EPS 15, IGF1R, MYC, PYCR1 and HNRNPA0 were downregulated, and the expressions of several classical factors involved in the growth and apoptosis of cancer cells were also decreased.

Conclusions: We found that SRBD1 were specifically expressed in non-small cell lung cancer tissue. Silencing of SRBD1 inhibits cell growth and promotes cell apoptosis in non-small cell lung cancer cells, and suppresses tumorigenesis in vivo, suggesting that SRBD1 may be a new diagnostic indicator and therapeutic target of non-small cell lung cancer.

摘要：

背景:研究S1 RNA結(jié)合域1 (SRBD1)在非小細(xì)胞肺癌組織中的表達(dá)及SRBD1沉默對(duì)人非小細(xì)胞肺癌細(xì)胞生物學(xué)行為的影響，探討SRBD1功能在人非小細(xì)胞肺癌細(xì)胞中的分子機(jī)制。

方法:采用免疫染色法和RT-PCR檢測(cè)SRBD1在人非小細(xì)胞肺癌組織和細(xì)胞系中的表達(dá)?；瘜W(xué)合成SRBD1的shrna，用慢病毒轉(zhuǎn)染A549和NCI-H1299細(xì)胞。通過(guò)細(xì)胞計(jì)數(shù)、MTT和克隆形成檢測(cè)細(xì)胞增殖。流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡。通過(guò)細(xì)胞注射BALB/c胸腺裸鼠評(píng)價(jià)細(xì)胞的致瘤性。采用基因芯片分析A549細(xì)胞的基因組變化。采用Ingenuity Pathway Analysis分析潛在的經(jīng)典信號(hào)通路、上游調(diào)控因子和基因相互作用網(wǎng)絡(luò)，并采用western blot分析進(jìn)行驗(yàn)證。

結(jié)果:SRBD1在人鱗狀細(xì)胞癌中特異表達(dá)，在肺癌細(xì)胞系NCI-H1299、A549和NCI-H1975中高表達(dá)。SRBD1定向shrna (shSRBD1)可有效降低A549和NCI-H1299細(xì)胞中SRBD1的表達(dá)。在裸鼠模型中，SRBD1沉默抑制非小細(xì)胞肺癌細(xì)胞增殖，促進(jìn)細(xì)胞凋亡，抑制腫瘤發(fā)生。此外，研究人員發(fā)現(xiàn)沉默SRBD1表達(dá)可導(dǎo)致A549細(xì)胞中基因表達(dá)的顯著變化。此外，shSRBD1組EPS 15、IGF1R、MYC、PYCR1和HNRNPA0蛋白水平下調(diào)，參與癌細(xì)胞生長(zhǎng)和凋亡的幾個(gè)經(jīng)典因子的表達(dá)也下降。

結(jié)論:研究人員發(fā)現(xiàn)SRBD1在非小細(xì)胞肺癌組織中特異性表達(dá)。沉默SRBD1可抑制非小細(xì)胞肺癌細(xì)胞生長(zhǎng)，促進(jìn)細(xì)胞凋亡，抑制體內(nèi)腫瘤發(fā)生，提示SRBD1可能成為非小細(xì)胞肺癌新的診斷指標(biāo)和治療靶點(diǎn)。

該論文中，人肺癌細(xì)胞系A549、NCI-H1299、NCI-H1975和95-D體外培養(yǎng)是使用Ausbian特級(jí)胎牛血清完成的。欲了解或購(gòu)買Ausbian特級(jí)胎牛血清可以聯(lián)系北京締一生物400-166-8600.