局部晚期非小细胞肺癌的治疗结果及预后相关microRNA研究

发布时间：2018-04-25 16:05

本文选题：局部晚期 + 非小细胞肺癌　；参考：《北京协和医学院》2017年博士论文

【摘要】：第一部分:局部晚期非小细胞肺癌同步放化疗的治疗结果目的:评价局部晚期非小细胞肺癌(non-small-cell lung cancer,NSCLC)患者同步放化疗的疗效和不良反应。方法:回顾性分析2001年1月至2010年12月间初治接受同步放化疗的251例局部晚期NSCLC患者的临床资料,其中IIIa期76例,IIIb期175例。放疗中位剂量为 60 Gy,其中调强放疗(intensity modulated radiation therapy,IMRT)174 例,三维适形放疗(three dimensional-conformal radiotherapy,3D-CRT)51 例,常规放疗 26例。全组患者接受EP方案(依托泊苷加顺铂)化疗112例,接受PC方案(紫杉醇加卡铂)化疗99例,接受拓扑替康单药化疗18例,接受其他方案化疗22例。分析同步放化疗的疗效及不良反应。结果:全组有244例患者可评价近期疗效,其中完全缓解(complete response,CR)6 例(2.5%),部分缓解(partial response,PR)183 例(75.0%),疾病稳定(stable disease,SD)42 例(17.2%),疾病进展(progressive disease,PD)13 例(5.3%)。全组患者中位随访时间62个月,中位生存时间为21个月,其1、3、5年生存率分别为69.2%、31.2%和23.2%。全组患者中位无进展生存时间为10个月,其1、3、5年无进展生存率分别为40.9%、22.1%和17.7%。Ⅲa期和Ⅲb期NSCLC患者的5年生存率分别为29.2%和20.7%,差异无统计学意义(p=0.133)。全组有244例患者可评价治疗失败模式,其中61例(25.0%)出现局部区域进展,55例(22.5%)出现远处转移,77例(31.6%)出现局部区域进展+远处转移。≥3级放射性肺炎、放射性食管炎及白细胞减少的发生率分别为4.4%、11.2%和26.7%。结论:局部晚期NSCLC同步放化疗的疗效较好,不良反应患者可耐受。同步放化疗是不可手术的局部晚期NSCLC的标准治疗。第二部分:局部晚期非小细胞肺癌同步放化疗后的巩固化疗结果分析目的:对于局部晚期非小细胞肺癌(non-small-cell lung cancer,NSCLC)患者,同步放化疗后巩固化疗的作用尚存在争议。基于此,本研究旨在分析巩固化疗的疗效和毒性。方法:回顾性分析2001年1月至2010年12月在我院接受根治性同步放化疗的局部晚期NSCLC患者的病例资料。结果:203例患者中,113例(55.7%)接受了巩固化疗。中位巩固化疗周期数为3个,89.4%的患者完成了≥2周期的巩固化疗。巩固化疗组的总生存(overall survival,OS)显著优于非巩固化疗组(中位OS,27个月vs.16个月;5年OS,30.4%vs.22.5%;p=0.012)。巩固化疗组和非巩固化疗组的中位无进展生存(progression free survival,PFS)分别为12个月和9个月(p=0.291)。亚组分析显示男性(HR:0.63;95%Cl,0.44-0.90)、年龄60 岁(HR:0.63;95%CI,0.42-0.95)、非鳞状细胞癌(HR:0.44;95%CI,0.25-0.76)、疗前卡氏评分≥80(HR:0.67;95%CI,0.48-0.93)、Ⅲb 期(HR:0.64;95%CI,0.43-0.95)、同步放化疗后疗效为稳定(stable disease,SD)(HR:0.31;95%CI,0.14-0.65)和放疗剂量≥60 Gy(HR:0.69;95%CI,0.48-1.00)的患者能够从巩固化疗中显著获益。毒性方面,巩固化疗组≥3级的血液学毒性发生率更高,但是两组差异没有达到统计学意义(45.1%vs.34.4%;p=0.123)。结论:同步放化疗的基础上加以巩固化疗可能能够进一步延长局部晚期NSCLC患者的生存而并没有增加治疗相关毒性,特别是对于男性、年龄60岁、非鳞癌、疗前卡氏评分≥80、Ⅲb期、SD和放疗剂量≥60 Gy的患者。但该研究结果仍然需要大规模前瞻性研究的证实。第三部分:MicroRNA-29c靶向VEGFA抑制肺腺癌进展的机制研究目的:肺腺癌具有侵袭性强和预后异质性大的特点,目前其发生发展的机制尚未被阐明。越来越多的证据显示microRNAs(miRNAs)的异常调控与多种肿瘤的发生发展密切相关。本研究目的为寻找肺腺癌的预后相关miRNA并研究其影响肿瘤进展的机制。方法:对87例病理证实为IIIa-N2期肺腺癌的福尔马林固定石蜡包埋(formalin-fixed paraffin-embedded,FFPE)标本进行miRNA芯片检测以寻找预后相关miRNA。采用脂质体转染法瞬时转染miR-29c过表达和干扰质粒,MTS实验检测细胞增殖能力变化,Transwell实验检测细胞迁移/侵袭能力变化。生物信息学和双荧光素酶报告基因实验预测和验证miR-29c与靶基因的关系,将miR-29c过表达和抑制后通过qRT-PCR、ELISA和Western blot检测靶基因mRNA、分泌蛋白和细胞内的蛋白变化。利用人脐静脉内皮细胞(human umbilical vein endothelial cell,HUVEC)成管实验检测miRNA对血管生成的影响。通过免疫组化(Immunohistochemistry,IHC)检测患者FFPE标本中miR-29c与微血管密度(microvessel density,MVD)和靶基因表达的相关性。结果:miRNA芯片及生存分析结果显示miR-29c表达下调与患者的不良预后显著相关。在体外实验中miR-29c抑制细胞增殖、迁移和侵袭。生物信息学和双荧光素酶报告基因实验发现miR-29c能够直接结合血管内皮生长因子A(vascular endothelial growth factor A,VEGFA)的 3'-UTR 区域,qRT-PCR、ELISA 和 Western blot实验结果发现miR-29c能够在转录和翻译水平下调VEGFA的表达。体外实验中miR-29c能够抑制HUVEC成管。高表达VEGFA能够基本完全逆转miR-29c导致的细胞增殖、迁移/侵袭和血管生成能力下降。在患者FFPE标本中miR-29c与MVD和VEGFA的表达具有显著相关性。结论:miR-29c通过靶向VEGFA在肺腺癌的进展中发挥抑癌作用,miR-29c有望作为肺腺癌预后的分子标志物和有价值的药物作用靶点。
[Abstract]:Part 1: therapeutic results of synchronous radiotherapy for locally advanced non-small cell lung cancer: To evaluate the efficacy and adverse reactions of concurrent chemoradiotherapy in patients with locally advanced non small cell lung cancer (non-small-cell lung cancer, NSCLC). Methods: a retrospective analysis of 251 locally advanced NSCL treated with synchronous radiotherapy from January 2001 to December 2010 The clinical data of C patients were 76 cases of IIIa phase and 175 cases in IIIb phase. The median dose of radiotherapy was 60 Gy, of which 174 cases were intensity modulated radiation therapy (intensity modulated radiation therapy, IMRT), 51 cases of three-dimensional conformal radiotherapy (three dimensional-conformal radiotherapy, 51 cases) and 26 cases with conventional radiotherapy. 112 cases were treated with PC regimen (paclitaxel plus carboplatin) chemotherapy in 99 cases, topotecan chemotherapy 18 cases, and other regimen chemotherapy 22 cases. Analysis of the curative effect and adverse reaction of concurrent chemoradiotherapy. Results: 244 patients in the whole group can evaluate the short-term effect, including complete remission (complete response, CR) 6 cases (2.5%), partial relief (partial response PR) 183 cases (75%), disease stability (stable disease, SD) 42 cases (17.2%), disease progression (progressive disease, PD) 13 cases (5.3%). The whole group was followed up for 62 months with a median survival time of 21 months, and its 1,3,5 year survival rate was 69.2%, 31.2% and 23.2%. group patients had no progression free survival for 10 months, and their 1,3,5 years did not advance. The 5 year survival rates of 40.9%, 22.1%, 17.7%. III A and III B NSCLC patients were 29.2% and 20.7%, respectively, with no statistical significance (p=0.133). 244 patients in the whole group evaluated the treatment failure mode, of which 61 cases (25%) had local regional progress, 55 (22.5%) had distant metastasis and 77 (31.6%) had local regional progress. Spreading + distant metastasis. The incidence of radiation pneumonitis of more than 3 levels, radiation esophagitis and leukocyte reduction were 4.4%, 11.2% and 26.7%., respectively: local advanced NSCLC radiotherapy and chemotherapy were better, patients with adverse reactions were tolerable. Synchronous radiochemotherapy was the standard treatment of Unoperable locally advanced NSCLC. Second part: locally advanced non small fine Analysis of the outcome of consolidation chemotherapy after synchronous radiotherapy and chemotherapy for cell lung cancer: for patients with locally advanced non-small cell lung cancer (non-small-cell lung cancer, NSCLC), the role of chemotherapy after concurrent chemoradiotherapy is still controversial. Based on this, the purpose of this study was to analyze the efficacy and toxicity of consolidation chemotherapy. Methods: a retrospective analysis from January 2001 to 2010 1 Results: 113 cases (55.7%) received consolidation chemotherapy in 203 patients in February. 113 of the 203 patients received consolidation chemotherapy. The median consolidation chemotherapy cycle number was 3, and 89.4% of the patients completed the consolidation chemotherapy for more than 2 cycles. The total survival (overall survival, OS) in the consolidation chemotherapy group was significantly better than non curing. The treatment group (median OS, 27 months vs.16 months; 5 years OS, 30.4%vs.22.5%; p=0.012). The median progression free survival (progression free survival, PFS) in the consolidation and non consolidation chemotherapy group was 12 months and 9 months respectively (p=0.291). The subgroup analysis showed that the male (HR:0.63; 95%Cl, 60), the age 60 years old, non squamous cells Cancer (HR:0.44; 95%CI, 0.25-0.76), before treatment, KRL score was more than 80 (HR:0.67; 95%CI, 0.48-0.93), stage III B (HR:0.64; 95%CI, 0.43-0.95). The curative effect was stable after concurrent chemoradiotherapy (stable disease) and radiotherapy doses of more than 60. The incidence of hematological toxicity in the cure group was more than 3, but the two groups were not statistically significant (45.1%vs.34.4%; p=0.123). Conclusion: the consolidation chemotherapy based on concurrent chemo chemotherapy could further prolong the survival of locally advanced NSCLC patients and did not increase the treatment related toxicity, especially for men, age 6. 0 years old, non squamous cell carcinoma, pre therapy score of more than 80, stage III B, SD and radiation dose more than 60 Gy. But the results still need a large prospective study. The third part: the mechanism of MicroRNA-29c targeting VEGFA to inhibit the progression of lung adenocarcinoma: lung adenocarcinoma is characterized by strong invasion and prognosis heterogeneity, and it is currently occurring The mechanism of development has not been clarified. More and more evidence shows that abnormal regulation of microRNAs (miRNAs) is closely related to the occurrence and development of various tumors. The purpose of this study is to search for the prognosis related miRNA of lung adenocarcinoma and to study the mechanism of its impact on the progression of the tumor. Methods: 87 cases of formalin fixed stone confirmed as IIIa-N2 lung adenocarcinoma were confirmed by pathology. Formalin-fixed paraffin-embedded (FFPE) specimens were tested for miRNA chip detection to find the prognosis related miRNA. using liposome transfection for transient transfection of miR-29c overexpression and interference plasmids, MTS test to detect cell proliferation ability, Transwell test to detect cell migration / invasiveness. Bioinformatics and double luciferase The relationship between miR-29c and target gene was predicted and verified by the reporter gene experiment. The target gene mRNA, secretory protein and protein changes were detected by qRT-PCR, ELISA and Western blot after the overexpression and inhibition of miR-29c. The angiogenesis of human umbilical vein endothelial cells (human umbilical vein endothelial cell) was tested for angiogenesis. The correlation between miR-29c and microvascular density (microvessel density, MVD) and target gene expression in FFPE specimens of patients with FFPE was detected by Immunohistochemistry (IHC). Results: miRNA chip and survival analysis showed that the downregulation of miR-29c expression was significantly related to the poor prognosis of the patients. The miR-29c inhibition in vitro was fine. Cell proliferation, migration and invasion. Bioinformatics and double luciferase reporter gene experiments found that miR-29c can directly bind the 3'-UTR region of vascular endothelial growth factor A (vascular endothelial growth factor A, VEGFA), and the qRT-PCR, ELISA, and Western experiments found that it can down the transcription and translation levels MiR-29c can inhibit HUVEC tube in vitro. High expression of VEGFA can completely reverse miR-29c induced cell proliferation, migration / invasion and decrease of angiogenesis. The expression of miR-29c and MVD and VEGFA in FFPE specimens of patients has significant correlation. Conclusion: miR-29c through target to VEGFA in the progression of adenocarcinoma of the lung to play a tumor suppressor. MiR-29c is expected to serve as a molecular marker for lung adenocarcinoma prognosis and a valuable target for drug use.

【学位授予单位】：北京协和医学院
【学位级别】：博士
【学位授予年份】：2017
【分类号】：R734.2

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