| Objectives1.To investigate the application of modified left RLN LNs dissection in MEE.2.To investigate the metastatic rate of bilateral RLN LNs in patients with esophageal cancer and its influencing factors.3.Correlation between histone modification changes and clinical and pathological staging and prognosis in esophageal cancer4.Mechanism of H2Bub over-expression in the occurrence and development of esophageal cancer.Methods and Results:the study is divided into four parts.Part ?:In this paper,39 cases were operated by the modified left RLN LNs dissection,which was comparatively studied with the 75 cases by traditional method and 80 cases by the skeletonized method.The result showed:1.the left RLN LNs metastasis was 18.56%(Group A:12%,Group B:20%,Group C:28%).Metastasis rate of Group A was significantly lower than that of Group C,which was statistically significant(?2=4.658 P=0.031).2.Average number of the left RLN LNs dissected of Group A(1.59±0.84)was significantly lower that that of Group B(4.02± 1.96)and Group C(4.31 ±2.14),which was statistically significant(P<0.05).3.The incidence of hoarseness in Group C was 15.4%,which was lower than that of Group B(21.3%)and higher than Group A(13.3%),however,there was no statistical difference between the three groups(P>0.05).Part ?:The paper retrospectively analyzed the data of 137 cases by MIE,and preliminarily studied the influencing factors to RLN LNs metastasis.The result showed:1.the right RLN LNs metastasis rate was 21.89%and the left was 21.17%and bilateral was 8.76%.2.Univariate analysis showed that the right RLN LNs metastasis was related with the differentiation,T staging and vessel carcinoma embolus in patients with esophageal cancer(P<0.05),while the left RLN LNs metastasis was related with the differentiation and T staging(P<0.05).3.Multivariate analysis showed that tumor differentiation degree and vessel carcinoma embolus was related to the right RLN LNs metastasis,with OR value of 3.707 and 3.053 respectively,and tumor differentiation was related to the left RLN LNs metastasis,with OR value of 3.960.Part ?:With para-carcinoma tissues as the controls,histone expression of H3K9Ac,H3K27Me3,H2BS112GlcNAc and H2Bub in 75 cases with esophageal carcinoma was detected by immunohistochemical method,and some of histone modifications related to the esophageal cancer were picked out.The result showed:1.Strongly positive expression rate of H2Bub,H3K9ac and H2BS112GlcNAc protein were 84%,48%and 21.3%in the tissue specimen of esophageal cancer,and positive expression rate of H3K27m3 protein were 52%.Compared with the para-carcinoma tissues,H2Bub,H3K9ac,H2BS112GlcNAc and H3K27m3 level in carcinoma tissues were 64%,46.7%,20.9%and 26.7%,which was up-regulated.H2Bub was related to the tumor differentiation,which was statistically significant(?2=0.243 P=0.037).2.The median survival time(36 months)of H2Bub in G1 group was significantly lower than that in the G2 and G3 group(71.63±11.21,58.88±3.95 months),which was with statistical difference(P<0.05).The median survival time of H3K27m3 expression level up-regulating group was better than that of the stable group with significant difference,which was statistically significant(P<0.05),suggesting that prognosis would be better by up-regulating H3K27m3 expression level.Regarding the H2BS112GlcNAc expression in esophageal cancer tissues,the differences in median survival time between strongly positive group and positive group,up-regulated group and stable group was statistically significant(P<0.05),suggesting that prognosis of strongly positive and up-regulated H2BS112GlcNAc expression would be poor.3.H3K9ac expression was co-related with the expression of H2Bub and H2BS112GlcNA(P<0.05).Part ?:Identify the H2Bub in the third part,construct over-expressed RNF20 plasmid and knock out USP22siRNA.Choose HEEC and ECA109 as study object,apply CCK8 and cell scratch experiment etc.to study the influence of H2Bub over-expression on cell proliferation and invasion.And the animal model of subcutaneous tumor bearing in nude mice was verified.Result:1.The transfection effect of overexpressed RNF20 plasmid is poor,so we plan to conduct lentivirus packaging experiment,but due to the interruption of the fight against covid-19,we will further supplement this experiment in the future.2.Lentivirus-mediated RNA interference successfully silenced the expression of USP22 in ECA109 and HEEC and established a stable cell line of H2Bub overexpression.3.CCK8 growth curve showed that the cell proliferation rate of the two cell lines was significantly faster than that of the empty carrier group,the difference of which was statistically significant(P<0.05).4.The cell scratch experiment showed that,compared with the empty carrier group,the scratch spacing between the two cell lines was significantly reduced trend.5.The subcutaneous tumor-bearing animal model of nude mice showed that the tumor size of the USP22 siRNA group was significantly larger than that of the NCsiRNA group(P<0.05).Conclusion:1.The modified left RLN LNs dissection method can provide sufficient LNs numbers dissected and reduce the incidence of RLN paralysis.It is safe and reliable in clinical works,which should be widely promoted and applied.2.Tumor differentiation degree and vessel carcinoma embolus were risk factors for the right RLN LNs metastasis,and tumor differentiation degree was the risk factor for the left RLN LNs metastasis.3.Expression level of H2Bub,H3K9ac and H2BS112GlcNAc in esophageal cancer tissue specimens was significantly higher than that of para-tumor tissues.Compared with the para-carcinoma tissues,H2Bub,H3K9ac,H2BS112GlcNAc and H3K27m3 level in cancer tissues was up-regulated in different degrees.4.Prognosis of up-regulated H2Bub expression level was poor in the poorly differentiated group.Prognosis was poor for the strongly positive and up-regulated H2BS112GlcNAc expression.Prognosis was better by up-regulating the H3K27m3 expression level.5.CCK8 experiment result showed that H2Bub can promote the proliferation and invasion of ECA109 and HEEC cells.Moreover,the subcutaneous tumor bearing animal model in nude mice verified that H2Bub can promote tumor formation in esophageal carcinoma. |
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