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The Soft Agar Colony Formation Assay
Published on: October 27, 2014
Circ_0067934 promotes non-small cell lung cancer development by regulating miR-1182/KLF8 axis and activating
Min Zhao1, Wei Ma1, Chaonan Ma1
1Department of Respiratory and Critical Medicine, The Huaihe Hospital of Henan University, Kaifeng, Henan, China.
Abstract:
Non-small cell lung cancer (NSCLC) is the primary subtype of lung cancer with high mortality. Circular RNAs (circRNAs) play a crucial role in tumor development and progression. This study aimed to explore the function of circ_0067934 in NSCLC progression and its molecular basis. The levels of circ_0067934, miR-1182 and kruppel like factor 8 (KLF8) were measured by quantitative real-time polymerase chain reaction or western blot assay. Cell viability was detected by Cell Counting Kit-8 (CCK-8) assay. Cell migration and invasion were assessed by transwell assay. Cell apoptosis was monitored by flow cytometry. The protein levels of epithelial-to-mesenchymal transition (EMT)-related markers and Wnt/β-catenin pathway-related proteins were examined by western blot. Dual-luciferase reporter assay, RNA Immunoprecipitation (RIP) assay or RNA pull-down assay was performed to verify the interaction among circ_0067934, miR-1182 and KLF8. Xenograft assay was used to detect tumor growth in vivo. We found that circ_0067934 and KLF8 were up-regulated, while miR-1182 was down-regulated in NSCLC tissues and cells. Circ_0067934 knockdown blocked proliferation, migration, invasion and EMT and induced apoptosis in NSCLC cells. Circ_0067934 regulated NSCLC progression by sponging miR-1182. MiR-1182 targeted KLF8 to hinder NSCLC development. In addition, depletion of circ_0067934 restrained tumor growth in vivo. In conclusion, Circ_0067934 acted as a competing endogenous RNA to facilitate NSCLC progression by regulating the miR-1182/KLF8 axis and activating Wnt/β-catenin pathway.
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