关键词: 白细胞分化抗原14, shRNA载体构建, 胃癌细胞, 侵袭

Abstract:

Background and purpose: Cluster of differentiation antigen 14(CD14), a high-affinity receptor for the lipopolysaccharide (LPS), could identify the gram-negative bacteria, fungi and mycobacterium tuberculosis, and mediate the inflammatory response of infected body, which is the first step in bacteria induced signal transduction. Expression of CD14 often turned to be abnormal in the tumor tissue and cancer cells. Recent studies have observed that functional CD14 polymorphisms, especially in the promoter motifs, are associated with a higher risk of H.pylori-related gastric carcinoma, indicated that CD14 is closely associated with the development of gastric cancer. This study aimed to construct CD14-shRNA expression vector and gastric cancer cells with CD14 silencing and to discuss the influence of CD14 on the invasion ability of gastric cancer cells and to lay a experimental basis for the study of the pathogenesis of gastric cancer. Methods: Four CD14-shRNA sequences were synthesized and CD14-shRNA expression vector was constructed to transfect cells and gastric cancer cells with CD14 silencing were screened by G418. RT-PCR and Western blot was used to detect the CD14 mRNA and protein level. The invasion ability of gastric cancer cells was detected by Transwell chamber. Results: CD14-shRNA expression vector was successfully constructed and screened to obtain gastric cancer cells with CD14 silencing, of which the CD14 mRNA and protein silencing efficiency were 71.7% and 63.4% respectively. Compared with the control group, the invasion ability of CD14-shRNA group was decreased obviously with statistical differences (P<0.01). Conclusion: Gastric cancer cells with CD14 silencing are constructed successfully and the influence of CD14 on the invasion ability of gastric cancer cells is preliminarily confirmed.

Key words: CD14, Construction of shRNA vector, Gastric cancer cells, Invasion