S1 RNA結合域1的沉默抑制人非小細胞肺癌細胞生長并促進細胞凋亡

2019年1月，北京協和醫學院國家腫瘤中心/國家腫瘤臨床研究中心/中國醫學科學院腫瘤醫院放射腫瘤科；北京環興腫瘤醫院放射腫瘤科(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老師研究團隊在《Translational lung cancer research》上發表論文：

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

“S1 RNA結合域1的沉默抑制人非小細胞肺癌細胞生長并促進細胞凋亡”

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結合域1 (SRBD1)在非小細胞肺癌組織中的表達及SRBD1沉默對人非小細胞肺癌細胞生物學行為的影響，探討SRBD1功能在人非小細胞肺癌細胞中的分子機制。

方法:采用免疫染色法和RT-PCR檢測SRBD1在人非小細胞肺癌組織和細胞系中的表達。化學合成SRBD1的shrna，用慢病毒轉染A549和NCI-H1299細胞。通過細胞計數、MTT和克隆形成檢測細胞增殖。流式細胞術檢測細胞凋亡。通過細胞注射BALB/c胸腺裸鼠評價細胞的致瘤性。采用基因芯片分析A549細胞的基因組變化。采用Ingenuity Pathway Analysis分析潛在的經典信號通路、上游調控因子和基因相互作用網絡，并采用western blot分析進行驗證。

結果:SRBD1在人鱗狀細胞癌中特異表達，在肺癌細胞系NCI-H1299、A549和NCI-H1975中高表達。SRBD1定向shrna (shSRBD1)可有效降低A549和NCI-H1299細胞中SRBD1的表達。在裸鼠模型中，SRBD1沉默抑制非小細胞肺癌細胞增殖，促進細胞凋亡，抑制腫瘤發生。此外，研究人員發現沉默SRBD1表達可導致A549細胞中基因表達的顯著變化。此外，shSRBD1組EPS 15、IGF1R、MYC、PYCR1和HNRNPA0蛋白水平下調，參與癌細胞生長和凋亡的幾個經典因子的表達也下降。

結論:研究人員發現SRBD1在非小細胞肺癌組織中特異性表達。沉默SRBD1可抑制非小細胞肺癌細胞生長，促進細胞凋亡，抑制體內腫瘤發生，提示SRBD1可能成為非小細胞肺癌新的診斷指標和治療靶點。

該論文中，人肺癌細胞系A549、NCI-H1299、NCI-H1975和95-D體外培養是使用Ausbian特級胎牛血清完成的。欲了解或購買Ausbian特級胎牛血清可以聯系北京締一生物400-166-8600.