Outcome And Predictors Of Neoadjuvant Chemoradiotherapy For Resectable Esophageal Squamous Cell Carcinoma The Role Of Postoperative Radiotherapy In Combined Small Cell Lung Cancer

Part IIMRT-based Neoadjuvant Chemoradiotherapy for Thoracic Esophageal Squamous Cell Carcinoma(ESCC):an Retrospective Analysis on Outcome and PrognosisObjective:Neoadjuvant chemoradiotherapy followed by surgery is recommended for patients with resectable esophageal carcinoma.However,after reviewing the evidence,we found that few study adopted modern radiation technique of IMRT,which can deliver more accurate dose to the target and avoid the surrounding organs as well.This study aimed to analyze the outcome,as well as the prognostic factors of IMRT-based neoadjuvant chemoradiotherapy followed by surgery in patients with thoracic ESCC.Methods and materials:Between Jan.2009 to May 2016,patients with thoracic ESCC who received IMRT-based neoajuvant chemoradiotherapy and surgery in our hospital were retrospectively analyzed.The UICC 2002 system was used when staging before surgery,while the UICC 2010 system was used after surgery.The median total radiation dose was 40Gy using 6MX x-rays at 1.8-2Gy/f,5 days per week.Concurrent chemotherapy was administered with platinum combined with 5-FU or paclitaxel.Adverse events were graded according to the RTOG/EORTC system.Kaplan-Meier analyses were used to assess survival.The Logrank test was used to examine differences between different groups.Cox’s proportional hazards model was used for multivariate analysis.A two-sided P-value of<5%was considered statistically significant.Results:Eighty-four patients were enrolled.The median age was 57.5 years(range,28-71 years).Endoscopic ultrasonography was used in 73 patients(86.9%).The disease was clinical stage II and III in 16 and 68 patients,which accounted for 19.0%and 81.0%,respectively.The median total radiation dose was 40Gy.After surgery,twenty-eight(33.3%)patients achieved pCR and 89.3%got down-staging.The R0 rate was 96.4%.There were no Grade IV adverse events.Grade III marrow suppression and esophagitis were seen in 8(9.5%)and 3(3.6%)patients,respectively.The postoperative mortality rate was 3.6%(two died of anastomotic fistula and one died of bleeding),and only 13 patients(15.5%)experienced postoperative adverse events.The median follow-up was 21 months(range,2.7-75.2 months).The 1-,3-and 5-year OS was 90.3%,63.6%,49.2%,respectively,with a corresponding DFS of 70.1%,55.5%and 47.0%,respectively.The 1-,3-and 5-year LRFS was 78.8%,55.6%,46.7%,respectively,with a corresponding DMFS of 77.3%,60.0%and 50.8%,respectively.On univariate analysis,clinical stageⅡ(p=0.025),R0(p=0.025)down-staging(p=0.022),good tumor response to neoajuvant chemoradiotherapy(p=0.021),pN0(p=0.036)and T/NpCR(p=0.029)were favorable prognostic factors.Clinical stage was the independent prognostic factor(95%CI 0.051-0.940,HR 0.220,p=0.041).Conclusions:For patients with thoracic ESCC,IMRT-based neoadjuvant chemoradiotherapy followed by surgery can achieve satisfied survival outcome.The incidence of postoperative complications is low.Clinical stage,down-staging,tumor response and pN are prognostic factors.Moreover,clinical stage was the independent prognostic factor.Part IIBiological Function of MiR-H1 in Inhibiting Esophageal Squamous Cell Carcinoma Cell Proliferation,Migration and InvasionObjective:Esophageal carcinoma(EC)is mostly seen in China and about 50%ES occur in China.However,unlike western countries,the pathology of EC in China is mostly squamous cell carcinoma.Currently,the five-year survival rate of esophageal squamous cell carcinoma(ESCC)remains poor.Considering the specialties of EC in our country,to explore the effective molecular biomarkers is of great value.MiRNAs play an important role in tumor processes(containing ESCC).In our previous study,we found that the high expression of miR-H1 was related to good prognosis.Our study aims to the biological function and mechanism of miR-H1 in ESCC.Methods and materials:The expression level of miR-H1 in 7 kinds of human esophageal squamous cell lines and human esophageal epithelial cell(HEEC)was quantified by the real-time qRT-PCR method.Lipofectamine 2000 was used for transfection of miRNA and co-transfection.Lentiviruses was used to infect cells making miR-H1 stably highly expressed.The proliferation of the transfected cells was evaluated by Cell Titer 96 AQueous One Solution Cell Proliferation Assay(MTS).The migration and invasion of transfected cells were evaluated by transwell assay.The potential target genes of miR-H1 were predicted using TargetScan and MIRDB.Then we used real-time qRT-PCR method and Western blot to confirm the relationship between miR-H1 and predicted genes in mRNA and protein level,respectively.Luciferase-reporter assay was applied to further investigate the gene’s role as a true target for miR-H1.NU/NU mice,4-6 weeks old,were injected subcutaneously and intravenously to observe the tumor volumn and metastasis.Results:In most human esophageal squamous cell lines,such as YSE2,KYSE 150,KYSE 510,the expressions of miR-H1 were largely lower than HEEC.The miR-Hl expression levels were increased by miR-H1 mimic and decreased by miR-H1 inhibitor,respectively,comparing with the control.By miR-Hl mimic transfection of KYSE 30 and KYSE 150,miR-H1 negatively regulated proliferation.In both YES2 and KYSE 30,the migration and invasion capabilities were decreased in miR-H1 over-expression group and increased in miR-Hl low-expression group,suggesting a tumor suppressor status for miR-H1.Through biological predictions,FMR1 was one of the potential targets of miR-H1.MiR-H1 and FMR1 mRNA,as well as miR-H1 and FMRP(the protein encoded by FMR1)showed negative correlations.The luciferase activity of FMR1-3’UTR,but not their mutant counterparts,was decreased by mimic in comparison with the control.The compromised cell proliferation,migration and invasion induced by miR-H1 mimic transfection were rescued by transfection of FMR1 expression plasmid in both YSE2 and KYSE 30.In vivo,tumors induced by miR-H1 overexpressed KYSE 30 grew significantly slower and resulted in smaller tumor mass.Metastatic lung colonization could be inhibited by overexpression of miR-HI in KYSE 30.Conclusions:MiR-H1 is significantly associated with survials and down-regulated in human esophageal squamous cell lines.MiR-H1 acts as a tumor suppressor by inhibiting proliferation,migration and invasion.FMR1 is the target of this miRNA.MiR-H1 is a promising biomarker and overexpressing miR-H1 might offer a potential therapeutic target against metastasis in patients with ESCC.Part IIIThe Role of Postoperative Radiotherapy(PORT)in Combined Small Cell Lung Cancer(C-SCLC)Objective:Combined small cell lung cancer(C-SCLC)is defined as small cell lung cancer(SCLC)combined with an additional component that consists of any of the histological types of non-small cell lung cancer(NSCLC).For patients with either SCLC or NSCLC who undergo surgery,postoperative radiotherapy(PORT)plays an important role in improving the treatment results.But there is no study focusing on PORT for C-SCLC yet.This single institutional study aimed to elucidate the effectiveness of PORT on survival of C-SCLC patients and to identify the subgroup which may most likely benefit from PORT.Methods and materials:Between Jan.2004 to Dec.2012,patients with histologically diagnosed C-SCLC and having a completely surgical resection in CIH-CAMS were retrospectively analyzed.The overall survival(OS),disease free survival(DFS),loco-regional recurrence free survival(LRFS),and distant metastasis survival(DMFS)were calculated by Kaplan-Meier method.A two-sided P-value of<5%was considered statistically significant.Results:Totally 55 consecutive patients were enrolled.The median follow-up was 56.4 months.The median OS,DFS,LRFS and DMFS were 62.2 months,43.4 months,54.9 months and 43.4 months,respectively.The 1-,3-,and 5-year OS rates were 86.6%,62.6%and 50.2%,respectively.For patients in the postoperative radiotherapy(PORT)and non-PORT groups,the median OS were 54.9 months and 62.2 months,respectively.The 1-,3-and 5-year OS rates were 92.9%,69.6%and 48.8%in the PORT group,and 84.4%,60.1%and 50.4%in the non-PORT group,respectively(p=0.605).The differences of DFS,LRFS and DMFS between PORT and non-PORT groups were also not statistically signifcant(p=0.466,p=0.455 and p=0.544).But analyzing the benefts in subgroups,PORT signifcantly improved survivals in stage III or N2 patients and patients with more than 10%of metastatic lymph nodes.For the patients with stage III disease,the 5-year OS,DFS,LRFS and DMFS were 61.0%、53.3%、61.0%and 53.3%in the PORT group,which were statistically signifcantly higher than the corresponding rates of 13.4%、13.8%、13.7%and 13.6%in the non-PORT group(p=0.015,0.026,0.008,0.030).For the patients with N2 stage,all survivals of the PORT group were also statistically signifcantly higher than non-PORT group(5y-OS:72.9%vs 11.1%,p=0.018;5y-DFS:62.5%vs 11.4%,p=0.032;5y-LRFS:72.9%vs 11.4%,p=0.008;5y-DMFS:62.5%vs 11.1%,p=0.042).Patients with more than 10%of metastatic lymph nodes could get a signifcant beneft survivals by receiving PORT(p=0.033,0.030,0.025,0.031).Having a systematic dissection of more than 17 lymph nodes was a subset which could get better OS and LRFS by receiving PORT(p=0.045,0.048),but not DFS and DMFS(p=0.109,0.121).Conclusions:PORT can signifcantly improve the survival of C-SCLC patients with resected pathological pN2 stage.For the patients with a large percent of metastatic lymph nodes,PORT can also improve survivals.