| BackgroundEsophageal cancer is one of the most common gastrointestinal malignancies these years the global incidence and mortality of it remains highly. the mortality of it in China is 15.15/10 ten thousand, 5-year survival rate is only 5%-10%, among which more than 95 percent are esophageal squamous cell carcinomas(ESCC). The early invasion and metastasis are the main factor which lead people’s death. and its mechanism is still unclear, it involves the interaction between tumor cells and host, it also associated with a variety of gene and protein expression.Heat shock proteins(HSPs) are a set of highly evolutionarily conserved proteins which could be induced and synthesized by various stimulations. Such as tumor, anoxia and other chemical stimulus. HSPs is also involved in the occurrence, development, immunity, therapy and prognosis of tumor. In recent years, a large number of studies have shown that HSP27 and HSP32, involved in a wide variety of tumor development, however, the role of HSP27 and HSP32 in the process of the invasion and metastasis of esophageal cancer remains unknown.ObjectiveTo research the significance of HSP27 and HSP32 in the invasion and metastasis of human esophageal squamous cell carcinoma(ESCC).Methods1. Study and research HSP32 and HSP27 expression levels in ESCC tissues and analyze the relationships with various clinicopathological features. 2. Using scratch experiment to distinguish the potential difference in vitro detection of esophageal squamous carcinoma height transfer cell subline transfer. 3. RT-PCR and western blot were done to test the changes of HSP32 and HSP27 in non-metastatic cells and high-metastatic ESCC cells. Analysis the influence of HSP32 and HSP27 in migration and invasion of esophageal squamous cell carcinomas. 4. Using lentiviral transfection technique expression promote the expression of HSP27 in highly metastatic sublines, via the experiments in vitro Transwell, cell scratch assay and nude mice,in vivo experiments to observe the changes of HSP27 raised after the invasion and metastasis of esophageal squamous cell carcinoma.Results1. Hps32 shows high expression in esophageal squamous cell carcinoma tissues the positive expressing rate and the degree of tumor differentiation(P=0.039), the infiltration degree of T staging(P=0.020), lymph node metastasis(P=0.017), lymphovascular invasion(P=0.037), distant metastasis(P=0.038) all of them have significant correlation; The decline of HSP27 in esophagus squamous carcinoma and metastatic cells system its expression is closely correlated with tumor differentiation(P=0.023), and TNM stage(P=0.013), lymph node metastasis(P=0.020) and distant metastasis(P=0.017) but it has no significant correlation with the gender(P=0.752) and the age of patients(P=0.771). 2. EC9706-H, EC109-H cell line invasion and metastasis ability of high; EC9706-L, EC109-L cell line with low invasion and metastasis. 3. HSP32 has a higher expression in two pairs of high metastatic cell subline thanin two pairs of low metastatic cell subline. On the contrary HSP27 with low metastatic of tow cell subline level show higher expression than tow tow cell subline level. 4. After infected the slow virus, it will raise the expression level of high cell subline of HSP27, Scratch assays and Transwell assyas both show that ability of HSP27 inmetastasis and invasion decline in ESCC cells. 5. Metastasis in nude mice experiment confirmed that the up regulation of HSP27 expression in high transfer will decline the invasion and metastasis ability of esophageal squamous carcinoma cell in vivo.Conclusion1. HSP32、HSP27 is closely related to the metastasis and invasion of ESCC, but different protein play different roles. The expression level of HSP32 was positively associated with esophageal carcinoma lymph; The expression level of HSP27 was negatively associated with esophageal carcinoma lymph. 2. Up-regulating HSP27 expression can restrain the ability of invasion and metastasi of sesophageal cancer cell and EC9706-H in vivo and in vitro. |
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