Clinical And Basic Research Of Micro MicroRNA-135b In Promoting The Progression Of Non-Small Cell Lung Cancer

Part 1Background: Non-small cell lung cancer(NSCLC)is one of the most commonly diagnosed cancers worldwide but with limited therapy efficacy.Uncovering the underlying pathological and molecular changes as well as mechanisms will improve the treatment.Dysregulated microRNAs play important roles in the initiation and progression of a wide range of malignancies,including NSCLC.Here,our research aims to investigate the biological functions of miR-135b in NSCLC.Methods: Public database mining,in situ hybridization and quantitative real-time PCR(qRT-PCR)were conducted to investigate the expression of miR-135b in NSCLC.Cell Counting Kit-8(CCK-8),colony formation,TNUEL,tube formation and transwell assays were performed to analyze the effects of miR-135b on cell growth and proliferation,apoptosis,migration,invasion and angiogenesis respectively in vitro.A xenograft tumor model was established to validate the effects of miR-135b on NSCLC progression in vivo.Luciferase reporter assays,western blotting were performed to identify the underlying mechanisms.Results: The level of miR-135b was significantly upregulated in NSCLC cell lines and tissues compared to the normal controls.NSCLC patients with higher expression of miR-135b got a poorer prognosis of survival.Overexpression of miR-135b facilitated,while inhibition of miR-135b impaired,cell proliferation,invasion,migration,survival and angiogenesis of NSCLC cells in vitro and in vivo.Mechanistic study revealed that miR-135b directly targeted the 3’-untranslated region(UTR)of CYLD m RNA to inhibit the expression of CYLD and further activate NF-B signaling and its downstream genes.Conclusion: We proved that miR-135b is tumor promoting in NSCLC.MiR-135b promotes tumor aggressiveness and progression in NSCLC through CYLD/NF-κB signaling.miR-135b can serve as a potential target for treatment of NSCLC patients.Part 2Background: Late cardiotoxicity related to radiotherapy(RT)in breast cancer and Hodgkin lymphoma has been well reported.However,the relatively higher cardiac dose exposure for esophageal cancer(EC)may result in earlier onset of cardiac diseases.In this report,we examined the incidence,onset,and long-term survival outcomes of high-grade cardiac events after RT in a large cohort of EC patients.Patients and Methods:Between March 2005 and August 2017,479 patients with EC from a prospectively maintained institutional database were analyzed.All patients were treated with either intensity modulated RT or proton beam therapy(PBT),either pre-operatively or definitively.We focused on any grade 3 or higher(G3+)cardiac events according to the Common Terminology Criteria for Adverse Events(CTCAE),version 5.0.Results: G3+ cardiac events occurred in 18% of patients at a median of 7 months with a median follow-up time of 76 months.Pre-existing cardiac disease(P=0.001)and radiation modality(P=0.027)were significantly associated with G3+ cardiac events.Under multivariable analysis,mean heart dose,particularly < 15 Gy,was associated to reduced G3+ events.Furthermore,G3+ cardiac events were associated with worse overall survival(P=0.041).Conclusion: Severe cardiac events were relatively common with early onset in EC patients after radiotherapy,especially those with pre-existing cardiac disease and higher radiation doses to the heart.Optimal treatment approaches should be taken to reduce cumulative doses to the heart,especially for patients with pre-existing cardiac disease.Part 3Purpose:Lymphocytes play a crucial role in antitumor immunity and they are also vulnerable targets during chemoradiotherapy(CRT).This study aimed to evaluate the impacts of different radiotherapy modalities on severe lymphopenia in a randomized trial involving patients with locally advanced esophageal cancer(EC).Patients and Materials : Between April 2012 and March 2019,a phase IIB,single-institutional,open-label,nonblinded,randomized trial was conducted.145 patients were randomly assigned to intensity modulated photon RT(IMRT)or proton beam therapy(PBT),either definitively or pre-operatively.The primary end point was the development of grade 4 lymphopenia(G4L)during concurrent CRT graded in accordance with the Common Terminology Criteria for Adverse Events version 5.0.Results:In total,105 patients were evaluable for analysis,with a median follow-up of 46 months.Among them,44 patients(42%)experienced G4 L which developed at a median of 28 days after the start of concurrent CRT.In multivariable analysis,radiation modality(IMRT vs.PBT,hazard ratio [HR]=2.782,95%CI,1.395-5.547,P=0.004)and planning target volume(PTV)(per 100 m L,HR=1.165,95%CI,1.043-1.302,P=0.007)were significantly associated with G4 L.In addition,dosimetric analysis showed that the mean lung dose(MLD)had the most significant effect on the incidence of G4L(HR=1.194;95%CI,1.083-1.316;P=0.000).There was no significant correlation between G4 L and survival outcomes.Conclusion:G4L is relatively common in EC patients during concurrent CRT.Compared with IMRT,PBT may be a good way to reduce the risk of severe,treatment-related lymphopenia with its dosimetric advantages.