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目的研究食管癌中是否存在螺杆菌感染及其与炎性信号通路的关系,探讨螺杆菌感染以及所产生的慢性炎症在人食管癌发生发展中的作用。方法随机收集食管癌标本37例和正常食管标本10例,采用聚合酶链反应(PCR)扩增螺杆菌16S rDNA,并进行基因测序及与幽门螺杆菌(Hp)同源性比较。阳性者再扩增Hp特异性毒力基因(cagA,babA2,vacA)。随后将Hp标准株NCTC11639和NCTC11637分别与食管鳞癌细胞株EC109共培养不同的时间,蛋白印迹(Western blot)检测炎性信号通路中P-IκB-α蛋白表达的变化。结果食管癌组织中螺杆菌16S rRNA基因的检出率为31.6%(12/37),正常人食管组织均无阳性表达。对阳性标本16S rRNA基因测序及同源性比较,食管癌组织中螺杆菌序列与Hp的16S rDNA序列同源性达99%~100%。12例阳性标本中分别有cagA阳性4例,babA2阳性8例,vacA阳性3例,阳性率分别为33%,67%,25%。EC109与Hp标准株共培养不同时间后,P-IκB-α显著升高。结论食管癌组织中存在螺杆菌感染,该螺杆菌可能为Hp或者其他亚种。螺杆菌感染导致食管粘膜慢性炎症在食管癌的发生发展中可能发挥重要的作用,其中P-IκB-α炎性信号通路的持续作用可能为分子机制之一。
Objective To investigate the relationship between Helicobacter infection and inflammatory signal pathways in esophageal cancer and to explore the role of Helicobacter infection and chronic inflammation in the development of human esophageal cancer. Methods 37 cases of esophageal cancer specimens and 10 cases of esophageal normal esophageal cancer specimens were randomly collected. The 16S rDNA of Helicobacter was amplified by polymerase chain reaction (PCR) and sequenced. The homology was compared with that of Helicobacter pylori (Hp). Hp-specific virulence genes (cagA, babA2, vacA) were amplified by positive amplification. Subsequently, Hp standard strains NCTC11639 and NCTC11637 were co-cultured with EC109 cells for different times. Western blot was used to detect the expression of P-IκB-α protein in the inflammatory signal pathway. Results The positive rate of 16S rRNA gene in Helicobacter pylori was 31.6% (12/37) in esophageal cancer tissues, but no in normal esophageal tissues. The positive 16S rRNA gene sequencing and homology comparison, esophageal cancer Helicobacter pylori sequence and Hp 16S rDNA sequence homology of 99% to 100%. Among the 12 positive samples, 4 were cagA positive, 8 were babA2 positive, and 3 were vacA positive. The positive rates were 33%, 67% and 25% respectively. After co-cultured with EC109 and Hp standard strains for different time, P-IκB-α was significantly increased. Conclusion There is Helicobacter infection in esophageal cancer tissues, which may be Hp or other subspecies. Helicobacter infection causes chronic inflammation of esophageal mucosa may play an important role in the occurrence and development of esophageal cancer. The continuing role of P-IκB-αin inflammatory signaling pathway may be one of the molecular mechanisms.