微创医学

目的探讨环磷酸腺苷反应元件结合蛋白转录辅激活因子3 （CRTC3）过表达对肺腺癌细胞NCI-H1975增殖和侵袭能力的影响。方法（1）收集15例肺腺癌患者的癌旁组织（距离癌组织>3 cm）和肺腺癌组织，采用实时荧光定量PCR检测组织中 CRTC3 mRNA 的表达水平。采用Western blotting检测正常人支气管上皮细胞（BEAS-2B）及非小细胞肺癌细胞系（H1299、NCI-H1975、HCC827、A549）的CRTC3蛋白表达水平。（2）将NCI-H1975 细胞分为空白对照组、阴性对照组、过表达组，空白对照组加入正常培养基，阴性对照组加入HBLV-ZsGreen-PURO（空载的慢病毒），过表达组加入LV-h-CRTC3-E/B（过表达载体），建立稳定过表达细胞株后，采用克隆形成实验检测细胞的增殖能力，采用Transwell 实验检测细胞的侵袭能力。结果（1）肺腺癌组织CRTC3 mRNA的相对表达水平低于癌旁组织（P<0.05）。与BEAS-2B及A549细胞比较，H1299细胞CRTC3蛋白相对表达水平升高，HCC827、NCI-H1975细胞CRTC3蛋白相对表达水平降低（P<0.05）。（2）与空白对照组和阴性对照组比较，过表达组NCI-H1975细胞的集落数量、侵袭数量减少（P<0.05）。结论与癌旁组织相比，肺腺癌患者的癌组织CRCT3 mRNA的表达水平降低，CRCT3过表达的NCI-H1975细胞株增殖及侵袭能力降低。

Objective To investigate the effect of overexpression of cyclic adenosine monophosphate response element-binding protein transcriptional coactivator 3 (CRCT3) on the proliferation and invasion abilities of the pulmonary adenocarcinoma cell NCI-H1975. Methods (1) Paracancerous tissues (at a distance of >3 cm from the cancerous tissues) and pilmonary adenocarcinoma tissues were collected from 15 patients of pulmonary adenocarcinoma. Quantitative real-time PCR was used to detect the expression level of CRTC3 mRNA in these tissues. Western blotting was applied to determine the expression level of CRTC3 protein in the normal human bronchail epithelial cell line (BEAS-2B) and non-small cell lung cancer cell lines (H1299, NCI-H1975, HCC827, A549). (2) NCI-H1975 cells were divided into the blank control group, the negative control group, and the overexpression group. The blank control group was added normal culture medium; the negative control group was transduced with HBLV-ZsGreen-PURO (empty lentivirus); the overexpression group was transduced with LV-h-CRTC3-E/B (overexpressing lentiviral vector). After establishing cell strains with stable overexpression, the colony formation assay was used to assess cell proliferation ability, and the Transwell assay was employed to evaluate cell invasion ability. Results (1) The relative expression level of CRTC3 mRNA in pulmonary adenocarcinoma tissues was lower than that in paracancerous tissues (P<0.05). Compared with BEAS-2B and A549 cells, the relative expression level of CRTC3 protein was increased in H1299 cells and decreased in HCC827 and NCI-H1975 cells (P<0.05). (2) Compared with the blank control group and the negative control group, the number of colonies and invasion were decreased in NCI-H1975 cells of the overexpression group (P<0.05). Conclusion Compared with paracancerous tissues, the expression level of CRCT3 mRNA in cancerous tissues of patients with pulmonary adenocarcinoma was decreased. Additionally, the proliferation and invasion abilities of the CRCT3 overexpressing NCI-H1975 cell strain was reduced.

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