Study On The Expression And Function Of HN1L In Esophageal And Gastric Junction Adenocarcinoma

Background:Adenocarcinoma of esophagogastric junction(AEG)is a special type of malignant tumor of digestive tract.In recent decades,the incidence of AEG continues to increase,which has received more and more clinical attention.Gastroesophageal reflux,smoking and obesity may be important causes of high incidence of AEG.Most patients with clinical symptoms are in the middle and advanced stage of cancer at the time of diagnosis,and the prognosis is very poor.With the development of science and technology,surgery,radiotherapy and chemotherapy have made great progress in recent years,but the 5-year survival rate of AEG patients is still very low.The pathogenesis of AEG is not clear.The gene HN1L(Hematologicalandneurologicalexpressed1 like),also known as L11,is located on the human chromosome 16p 13.3.A number of studies have shown that the expression of HN1L is increased in non-small cell lung cancer,liver cancer,breast cancer and other malignant tumors,and may promote malignant proliferation and metastasis of tumors through different signal pathways.But at present,the relationship between HN1L and AEG is still a mystery.In this study,38 AEG patients and AEG cell lines were used to detect the expression of HN1L in AEG tissues,and to explore the effects of HN1L on the proliferation,migration,invasion and apoptosis of AEG-related cell lines,so as to provide theoretical support for further study of its molecular mechanism.Methods:1.Immunohistochemical staining was used to detect the expression of HN1L in cancer and adjacent tissues of 38 patients with AEG.2.The clinicopathological features of 38 patients were analyzed,and the relationship between clinicopathological features and the expression level of HN1L was analyzed.3.The effective experimental cell lines were screened by real-time fluorescence quantitative PCR(qRT-PCR)to construct the AEG cell line with low HN1L.Knock-down HN1L interference lentivirus and control lentivirus were constructed and packaged by Shanghai Jikai Company and transfected into AGS and HGC-27 cell lines respectively.4.The effect of knocking down HN1L on the proliferation of AEG cells was detected by Celigo real-time monitoring technology,clone formation test and MTT experiment.5.Transwell assay was used to detect the effect of knocking down HN1L on the migration and invasion of AEG cells.6.The effect of knocking down HN1L on apoptosis of AEG cells was detected by flow cytometry.Result:1.The expression level of HN1L in AEG carcinoma was higher than that in paracancerous tissues.There was a significant difference in the expression of HN1L between AEG carcinoma tissue(n=38)and corresponding paracancerous tissue(n=38)(P<0.001).2.There was no statistical significance in gender,age,tumor length or Siewert classification of patients in high HN1L expression group(score ≥ 3.4)and low HN1L expression group(score<3.4)(P>0.05).The expression level of HN1L was significantly correlated with TNM stage(P=0.013)and lymph node metastasis(P=0.03)in AEG patients.3.QRT-PCR detection showed that the expression level of HN1L in AGS and HGC-27 cell lines was higher than that in MKN-45 and MGC80-3 cell lines.Therefore,AGS and HGC-27 cell lines were selected as experimental cells.AGS and HGC-27 cell lines stably knocking down HN1L were successfully constructed.4.Knocking down HN1L can inhibit the proliferation of AEG cells in vitro.After continuous detection by Celigo instrument,the cell growth curve showed that the proliferation rate of AEG cells decreased significantly after knocking down HN1L,clone formation assay showed that the number of clones of AEG cells decreased significantly after knocking down HN1L,and MTT assay showed that the proliferation ability of AEG cells decreased significantly after knocking down HN1L.5.Knocking down HN1L can inhibit the migration and invasion of AEG cells.Transwell migration assay showed that the number of AEG cells passing through polycarbonate membrane decreased significantly after knocking down HN1L.Further Transwell invasion assay showed that the number of cells passing through polycarbonate membrane of AEG cells decreased significantly after knocking down HN1L.6.Knocking down HN1L can promote the apoptosis of AEG cells.The results of flow cytometry showed that the apoptosis rate of AEG cells increased significantly after knocking down HN1L.Conclusion:1.The expression level of HN1L in AEG tissues was higher than that in paracancerous tissues.2.The expression of HN1L was significantly correlated with TNM stage and lymph node metastasis in patients with AEG.3.Knocking down HN1L can inhibit the proliferation,migration and invasion of AEG cells,and promote the apoptosis of AEG cells.This study shows that HN1L may be the oncogenic gene of AEG and may become a new target for the diagnosis and treatment of AEG.