连翘苷调控LINC00342影响胃癌细胞增殖、迁移和侵袭及炎症因子表达的实验研究

Forsythin regulates cell proliferation,migration,and invasion and expression of inflammatory factors in gastric cancer cells by regulating LINC00342

背景连翘苷是中药连翘的主要活性成分之一,具有抗肝癌、肺癌和肾癌的作用,但其能否影响胃癌细胞的恶性行为还未知.长基因间非编码RNA 00342(LINC00342)是一种非小细胞肺癌和结直肠癌中表达增加的长链非编码RNA(lncRNA),其作为促癌基因促进这些肿瘤细胞的恶性行为,进而促进肿瘤的发展进程.本研究假设连翘苷可通过抑制LINC00342发挥抗胃癌作用.目的探讨连翘苷对胃癌细胞增殖、迁移、侵袭和炎症因子表达的影响及可能机制.方法体外培养胃癌HGC-27细胞,用不同剂量(5、10、20μmol/L)连翘苷干预24 h后,CCK-8和克隆形成实验检测检测细胞增殖,Transwell检测细胞迁移和侵袭,蛋白质印迹法检测细胞中E-cadherin和N-cadherin蛋白表达,试剂盒检测细胞培养上清液中炎症因子TNF-α和IL-6表达,qRT-PCR法检测细胞中LINC00342表达.收集51例胃癌组织和癌旁组织,qRT-PCR法检测组织中LINC00342表达.转染LINC00342小干扰RNA或过表达载体至HGC-27细胞,上述相同方法观察LINC00342对HGC-27细胞增殖、迁移、侵袭及炎症因子表达的影响.结果与对照组比较,HGC-27细胞经不同剂量连翘苷干预后,细胞增殖抑制率、E-cadherin蛋白表达量均升高(P<0.05),细胞克隆数、迁移数、侵袭数及细胞中N-cadherin蛋白和LINC00342的表达量、细胞培养上清液中TNF-α和IL-6表达均降低(P<0.05).胃癌组织中LINC00342的表达量为显著高于癌旁组织(P<0.05).敲减LINC00342后,HGC-27细胞增殖抑制率、E-cadherin蛋白表达量均升高(P<0.05),细胞克隆数、迁移数、侵袭数、细胞中N-cadherin蛋白表达、细胞培养上清液中TNF-α和IL-6表达均降低(P<0.05).过表达LINC00342逆转了连翘苷对HGC-27细胞增殖、迁移、侵袭和炎症因子表达的影响.结论连翘苷可抑制胃癌HGC-27细胞增殖、迁移和侵袭及炎症因子TNF-α和IL-6表达,其作用机制可能与下调细胞中LINC00342表达有关.

BACKGROUND Forsythin is one of the main active components of the traditional Chinese medicine forsythia.It has anti-liver cancer,lung cancer,and kidney cancer effects,but whether it affects the malignant behavior of gastric cancer cells is still unknown.Long intergenic non-coding RNA 00342(LINC00342)is a long non-coding RNA with increased expression in small cell lung cancer and colorectal cancer,which acts as an oncogene to promote the malignant behavior of these tumor cells,thereby promoting the development of tumors.We hypothesized that forsythin has an anti-gastric cancer effect by inhibiting LINC00342.AIM To investigate the effect of forsythin on the cell proliferation,migration,and invasion and the expression of inflammatory factors in gastric cancer cells and the possible mechanism involved.METHODS Gastric cancer cells(HGC-27)were cultured in vitro and treated with different doses(5,10,and 20μmol/L)of forsythin for 24 h.Cell proliferation was detected by CCK-8 and colony formation assays,and cell migration and invasion were detected by Transwell assay.The protein expression of E-cadherin and N-cadherin in cells was detected by Western blot,and the expression of inflammatory factors TNF-αand IL-6 in cell culture supernatant was detected with commercial kits.qRT-PCR was used to detect the expression of LINC00342 in cells.Fifty-one cases of gastric cancer tissues and adjacent tissues were collected,and qRT-PCR was used to detect the expression of LINC00342 in the tissues.LINC00342 small interfering RNA or overexpression vector was transfected into HGC-27 cells,and the effects of LINC00342 on HGC-27 cell proliferation,migration,and invasion and the expression of inflammatory factors were explored.RESULTS Compared with the control group,the proliferation inhibition rate and the protein expression of E-cadherin in HGC-27 cells treated with different doses of forsythin were increased(P<0.05),but the number of cell clones,migration and invasion,the expression of N-cadherin protein and LINC00342 in the cells,and the expression of TNF-αand IL-6 in the cell culture supernatant were all decreased(P<0.05).The expression of LINC00342 in gastric cancer tissue was significantly higher than that in adjacent tissues(P<0.05).After knocking down LINC00342,the proliferation inhibition rate and the protein expression of E-cadherin in HGC-27 cells treated with different doses of forsythin were increased(P<0.05),but the number of cell clones,migration and invasion,the protein expression of N-cadherin in the cells,and the expression of TNF-αand IL-6 in the cell culture supernatant were decreased(P<0.05).Overexpression of LINC00342 reversed the effects of forsythin on the cell proliferation,migration,and invasion and the expression of inflammatory factors in HGC-27 cells.CONCLUSION Forsythin may inhibit the proliferation,migration,and invasion of gastric cancer HGC-27 cells and the expression of inflammatory factors TNF-αand IL-6 via mechanisms that may be related to the down-regulation of the expression of LINC00342.