Objective To study the inhibition effects of various gastrin-shNAs on gastrin expression in gastric cancer cell line BGC-823. Methods Four nucleotide sequences of shRNA were designed corresponding to various sites of gastrin gene.Four shRNAs were synthesized by in vitro transcription and transfected into gastric cancer cell line BGC-823 at the final concentration of 10nmol/L,20nmol/L,40nmol/L and 80nmol/L respectively.In situ hybridization and immunohistochemistry techniques were applied to investigate the inhibition of gastrin expression and screen the most effective shRNA.The inhibitory effect on gastrin mRNA of screened shRNA was further identified by RT-PCR.MTT assay was used to determine the inhibitory effect of 4 shRNAs at various final concentrations on the growth of BGC-823 cells. Results The gastrin mRNA and protein exression were suppressed distinctly 24,48,and 72hours after transfection,and exhibited time-and concentration-dependent tendency.The highest suppression efficiency on both mRNA(54.27?0.042)% and protein(41.69?0.038)% level occurred 72 hours later in the cells transfected with shRNAs.The RT-PCR result showed that the inhibitory ratio of shRNA3 on gastrin mRNA of BGC-823 was 48.1%.MTT displayed a proliferative inhibition of the BGC-823 cells after transfection of shRNAs with a concentration-denpendent tendency except the shRNA4 treated cells.Conclusion Four gastrin-shRNAs showed a significant inhibition effect on gastrin expression of gastric cancer cell BGC-823 on mRNA and protein level.shRNAs might be the most effective gastrin-shRNA.Inhibited gastrin expression by shRNAs resulted in a significant decrease of proliferative ability of BGC-823 cells.

Objective To analyze copy number variance (CNV) in whole genome by using gene chip technology, and to screen the radiosensitivity associated genes on esophageal squamous cell carcinoma (ESCC). Methods The patients with ESCC who received radiotherapy alone in Anyang Tumor Hospital from December 2013 to August 2016 were selected, and biopsy paraffin samples were preserved in the center of pathology. The patients were divided into radiosensitivity group (group S) and radio-resistance group (group R). DNA was extracted from these paraffin samples in both groups. Whole human genome CNV was detected by using genechip from OncoScan Array platform designed by Affymetrix company, and the differences of gene segments were screened in the two groups. Results Nineteen samples of ESCC patients were collected to extract DNA in this study. To balance pair analysis in the two groups, 10 samples were selected from the qualified patients, including 5 cases in group S and 5 cases in group R respectively. There were no statistical differences in gender, age, lesion site, lesion length, radiation dose of the two groups (all P> 0.05). Loss of heterozygosity (LOH) was the main type of CNV. The analysis results showed that LOH in q24.32-q24.33 of chromosome 10 and LOH in q21.2-q21.31 of chromosome 18 had high frequencies (100 ％) in group R, however, none were detected in group S. LOH in q27-q28.1 of chromosome 4 had a high frequency (80％) in group S , however, none were detected in group R. Conclusion LOH in 10q/18q is related to radio-resistance in ESCC, and LOH in 4p is associated with radiosensitivity in ESCC.

OBJECTIVETo investigate the predictive value of P53, Ki-67, HER2 protein detection in neoadjuvant chemotherapy for adenocarcinoma of gastroesophageal junction (AGEJ).

METHODSPreoperative biopsy specimens and clinical data of 72 patients of locally advanced Siewert II AGEJ between June 2010 and December 2013 were reviewed. All the patients received SOX scheme neoadjuvant chemotherapy, and were divided into effective group (complete response plus partial response) and ineffective group (stable disease plus progressive disease). Expressions of above 3 proteins were detected by immunohistochemistry in all the patients before neoadjuvant chemotherapy. The relationship between various proteins and efficacy of chemotherapy was analyzed by univariate and logistic multivariate regression analyses.

RESULTSAll the 72 patients successfully completed 2 cycles of SOX neoadjuvant chemotherapy, among them, 5 cases (6.9%) with complete response, 30 cases (41.7%) with partial response, 31 cases (43.1%) with stable disease, 6 cases (8.3%) with progressive disease, including 35 cases in effective group and 37 cases in ineffective group. Compared with ineffective group, the positive expression rate of P53 was significantly reduced (25.0% vs. 45.9%, P=0.020), and that of Ki-67 significantly increased (77.1% vs. 43.2%, P=0.003), however, there was no significant difference in the expression rate of HER2 between the two groups (P>0.05). Multivariate analysis showed that Ki-67 was the independent predictive factor for the efficacy of neoadjuvant chemotherapy (P=0.015). Spearman rank correlation showed that Ki-67 expression was positively correlated with HER2 expression (r=0.259, P=0.028), but P53 expression was not correlated with Ki-67 or HER2 (r=0.140, 0.042, P=0.240, 0.725, respectively).

CONCLUSIONSSOX neoadjuvant chemotherapy is safe and effective for AGEJ, especially for patients with depressed expression of P53 and elevated expression of Ki-67, which both may be used as reference for the prediction of chemotherapy efficacy. There is no correlation between P53 and Ki67 proteins, so combined detection may improve the predictive value.

Adenocarcinoma ; diagnosis ; drug therapy ; Esophageal Neoplasms ; diagnosis ; drug therapy ; Esophagogastric Junction ; pathology ; Humans ; Immunohistochemistry ; Ki-67 Antigen ; metabolism ; Neoadjuvant Therapy ; Receptor, ErbB-2 ; metabolism ; Remission Induction ; Tumor Suppressor Protein p53 ; metabolism