S1 RNA结合结构域1的沉默抑制人非小细胞肺癌细胞的生长并促进其凋亡。
Silence of S1 RNA binding domain 1 represses cell growth and promotes apoptosis in human non-small cell lung cancer cells.
作者信息
Zhang Tao, Cheng Guowei, Deng Lei, Yang Yin, Sun Li, Chen Ping, He Xiangling, Su Dan, Bi Nan, Qiu Bin
机构信息
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, Beijing 10021, China.
出版信息
Transl Lung Cancer Res. 2019 Dec;8(6):760-774. doi: 10.21037/tlcr.2019.10.10.
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 , suggesting that SRBD1 may be a new diagnostic indicator and therapeutic target of non-small cell lung cancer.
背景
研究S1 RNA结合结构域1（SRBD1）在非小细胞肺癌组织中的表达以及SRBD1沉默对人非小细胞肺癌细胞生物学行为的影响，并探讨SRBD1在人非小细胞肺癌细胞中的功能分子机制。
方法
通过免疫染色和RT-PCR检测SRBD1在人非小细胞肺癌组织和细胞系中的表达。化学合成SRBD1的短发夹RNA（shRNA），并通过慢病毒转染至A549和NCI-H1299细胞。通过细胞计数、MTT和克隆形成检测细胞增殖。通过流式细胞术检测细胞凋亡。通过将细胞注射到BALB/c裸鼠中评估致瘤性。采用基因芯片分析探索A549细胞中的基因组变化。通过Ingenuity Pathway Analysis分析潜在的经典信号通路、上游调节因子和基因相互作用网络，并通过蛋白质印迹分析进行验证。
结果
SRBD1在人鳞状细胞癌中特异性表达，在肺癌细胞系NCI-H1299、A549和NCI-H1975中高表达。SRBD1靶向shRNA（shSRBD1）有效降低了A549和NCI-H1299细胞中SRBD1的表达。SRBD1沉默抑制了非小细胞肺癌细胞的增殖，促进了细胞凋亡，并在裸鼠模型中抑制了肿瘤发生。此外，我们发现SRBD1表达沉默导致A549细胞中的基因表达发生显著变化。此外，在shSRBD1组中，EPS 15、IGF1R、MYC、PYCR1和HNRNPA0的蛋白水平下调，参与癌细胞生长和凋亡的几个经典因子的表达也降低。
结论
我们发现SRBD1在非小细胞肺癌组织中特异性表达。SRBD1沉默抑制非小细胞肺癌细胞的生长并促进细胞凋亡，抑制肿瘤发生，提示SRBD1可能是非小细胞肺癌的一种新的诊断指标和治疗靶点。
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