miR-214通过MAPK信号通路对非小细胞肺癌放疗敏感性的影响

目的：探讨miR-214通过MAPK信号通路对非小细胞肺癌放疗敏感性的影响。方法：体外培养构建放射抵抗性的肺癌H358细胞株（H358-RR）;qPCR实验检测H35和H358-RR细胞中miR-214的表达水平,qPCR检测miR-214-mimic在H358-RR中的转染效率及miR-214的表达情况;Western blot检测miR-214-mimic对ERK1、jnk和p38MAPK蛋白表达水平的影响;免疫荧光检测γ-H2AX聚集点的情况及对放射的敏感性;CCK-8实验检测H358-RR细胞株接受放射后活性的变化;单细胞凝胶电泳检测放射线照射后不同组肺癌细胞DNA损伤情况。结果：成功构建H358-RR放射抵抗细胞株,qPCR检测H358-RR细胞中miR-214的表达水平高于H358细胞,qPCR检测miR-214-mimic的转染效率良好,可以有效增加miR-214的表达水平,Western blot检测转染miR-214-mimic后ERK1、jnk和p38MAPK的表达水平相应升高;过表达miR-214之后,免疫荧光检测γ-H2AX聚集点的表达明显较少,DNA损伤较少;过表达miR-214后,H358-RR细胞接受放射后细胞活力降低水平减少;过表达miR-214后H358-RR抵抗放射线及对DNA损伤修复的能力增强。结论：miR-214通过影响MAPK信号通路调控非小细胞肺癌的DNA损伤及修复能力,影响其对放疗的敏感性。     

非小细胞肺癌患者放疗前后机体免疫功能变化的研究

目的：报道３０例非小细胞肺癌（ＮＳＣＬＣ）患者放疗前后免疫功能的变化。方法：采用双抗体夹心法和比浊法检测血清中ｓＩＬ－２Ｒ、免疫球蛋白（ＩｇＧ,ＩｇＡ,ＩｇＭ）含量,用单克隆抗体和流式细胞仪技术检测外周血中Ｔ细胞亚群,Ｂ细胞,ＮＫ细胞百分数;与健康人做对照。结果：放疗前ｓＩＬ－２Ｒ显著增高（Ｐ〈０．０１）,ＩｇＧ及ＩｇＭ６亦增高（Ｐ〈０．０５）,Ｂ细胞百分数低于（Ｐ〈０．０１）,ＣＤ３＾＋细胞降低     

Wip1在非小细胞肺癌组织及细胞中的表达

目的： 检测肺癌组织和3种肺癌细胞中Wip1的表达水平,探讨肺癌中Wip1的表达水平与其各种临床病理特征之间的关系。方法: 利用实时荧光定量PCR检测44例肺癌及相应正常肺组织中Wip1 mRNA的表达,分析Wip1 mRNA表达与各临床病理参数之间的关系;利用实时荧光定量PCR的方法检测人肺腺癌细胞A549、人鳞癌细胞NCI-1299、人大细胞肺癌细胞NCI-H460和人正常支气管上皮细胞HBE中Wip1 mRNA的表达量,进行相对定量分析。结果: 44例非小细胞肺癌及相应正常肺组织均有Wip1 mRNA的表达,其中17例肺癌中Wip1 mRNA高表达,占38.6%,两者表达差异显著(ratio=2.1644±1.3940,P＜0.05);分化程度低的肿瘤细胞Wip1 mRNA表达量显著高于分化程度高者（P<0.05）。3种肺癌细胞中的Wip1 mRNA表达量显著高于正常支气管上皮细胞,差异显著(均P＜0.05)。结论: Wip1mRNA在非小细胞肺癌中过表达,可能与肿瘤发生有关,有望成为非小细胞肺癌基因治疗的新靶点。Wip1 mRNA在非小细胞肺癌中的表达与肿瘤细胞分化程度有关,可能成为确定肿瘤恶性程度的分子生物学参考指标。     

非小细胞肺癌中P53表达的不同亚细胞定位及功能意义的研究

目的:探讨非小细胞肺癌(NSCLC)中不同亚细胞定位P53蛋白及其可能的功能意义。方法:以免疫组化及流式细胞分析术研究了43例NSCLC病人肿瘤组织P53表达定位与G₁/S检查点功能及DNA含量的关系。结果:发现P53蛋白不同的亚细胞定位与病人的病程密切相关,同时,在P53核表达定位的病人肿瘤组织细胞S期比例明显增多而G₀－G₁期比例明显较胞浆表达病人减少,G₂－M期比例及DNA指数在两组间无明显差异。结论:不同亚细胞定 位的P53蛋白具有不同的G₁/S检查点调节功能,胞浆P53蛋白仍有部分G₁/S检查点调控功能,因此出现较少的SPF及较多的G₀－G₁细胞,代表了恶性度较低的细胞亚群;而核表达阳性P53蛋白具有更差的G₁/S检查点调控功能,代表了恶性度较高的肿瘤亚群,这与肿瘤进展可能有关。     

非小细胞肺癌不同分割剂量三维适形放疗与常规放疗的比较

%(7/81).3组患者局部控制率及生存率曲线差异具有统计学意义(均P<0.05).放射损伤主要为放射性肺纤维化和放射性食管炎,CRT组发生率较高(P<0.05).结论 3DCRT对NSCLC有较好的疗效,放疗损伤较CRT低,尤以3DCRT Ⅰ组疗效较好,更具优势.     

非小细胞肺癌不同分割剂量三维适形放疗与常规放疗的比较

%(7/81).3组患者局部控制率及生存率曲线差异具有统计学意义(均P<0.05).放射损伤主要为放射性肺纤维化和放射性食管炎,CRT组发生率较高(P<0.05).结论 3DCRT对NSCLC有较好的疗效,放疗损伤较CRT低,尤以3DCRT Ⅰ组疗效较好,更具优势.     

Clinicopathological and prognostic significance of microRNA-107 in human non small cell lung cancer

Background: MicroRNAs (miRNAs) are small, non-coding RNAs which can function as oncogenes or tumor suppressor genes in human cancers. Researchers have found that the expression level of miR-107 was decreased in human non-small cell lung cancer (NSCLC) tissues and cell lines, however, its clinicopathological and prognostic significance in NSCLC has not been investigated. Methods: Quantitative real-time PCR (qRT-PCR) was used to analyze the expression of miR-107 in 137 pairs of fresh NSCLC and matched adjacent normal tissue specimens. The chi-square test and Fishers exact tests were used to examine the associations between miR-107 expression and the clinicopathological characters. The overall survival (OS) and progression-free survival (PFS) were analyzed by log-rank test, and survival curves were plotted according to Kaplan-Meier. Results: The expression level of miR-107 was significantly lower in tumor tissues than that in corresponding noncancerous tissues (0.4676±0.2078 vs. 1.000±0.3953, P<0.001). Low expression of miR-107 was found to significantly correlate with TNM stage (p=0.001), regional lymph node involvement (p=0.04), and tumor differentiation (p=0.003). Kaplan-Meier analysis with the log-rank test indicated that low miR-107 expression had a significant impact on OS (35.2% vs. 69.3%; P=0.008) and PFS (30.0% vs. 56.2%; P=0.029). In a multivariate Cox model, we found that miR-107 expression was an independent poor prognostic factor for both 5-year OS (HR=2.57, 95% CI: 1.88-10.28; P=0.007) and 5-year PFS (HR=3.08, 95% CI: 2.01-8.92; P=0.003). Conclusion: The expression of miR-107 was decreased in NSCLC. Low expression of miR-107 was significantly associated with tumor progression and decreased survival in patients with NSCLC, indicating that miR-107 may serve as a novel prognostic marker in NSCLC.     

人非小细胞肺癌中KGF mRNA的表达及意义

目的研究人非小细胞肺癌(non-small cell lung cancer,NSCLC)角化细胞生长因子(keratinocyte growth factor,KGF)mR-NA的表达,及其在NSCLC发生过程中肿瘤细胞与间质细胞间的相互作用。方法采用原位杂交和免疫组化法检测KGF mR-NA与Ki-67在50例NSCLC的表达,并与正常组织对照。结果KGF mRNA的表达除在NSCLC某些实质细胞内观察到外,主要见于NSCLC的纤维母细胞和血管平滑肌细胞胞质。肿瘤组织KGF mRNA表达的阳性率86%明显高于正常肺组织的24%(P<0·05)。有淋巴结转移者比无淋巴结转移者的表达更强,且与肺癌的分化相关,分化程度越低,KGF mRNA表达越强。在50例肺癌中Ki-67表达的分布与KGF mRNA相似。结论NSCLC存在KGF mRNA高表达。KGF可通过旁分泌、自分泌两种方式发挥作用。KGF可能与NSCLC的发生有一定的相关性。     

miR-183及MAPK1蛋白在非小细胞肺癌中的表达及其意义

目的探讨miR-183及丝裂原激活的蛋白激酶1（MAPKl）mRNA与蛋白在非小细胞肺癌（NSCLC）组织中的表达及意义。方法实时定量PCR方法检测NSCLC组织miR-183及MAPKlmRNA的表达水平,Westernblot方法检测NSCLC组织MAPKl蛋白表达;统计分析miR-183与MAPKl蛋白的相关性。结果miR-183在非小细胞肺癌组织中的表达量较癌旁组织明显降低（P〈0．01）,MAPKlmRNA与蛋白在癌组织中较癌旁组织表达明显增高（P〈0．05）;统计分析显示,miR-183表达与MAPKl表达呈显著地负相关关系。结论miR-183在NSCLC组织中低表达,与MAPKl表达呈显著负相关关系。     

Expression of epithelial and neural antigens in small cell and non small cell lung carcinoma

Seventy-one lung carcinomas from 66 different patients were stained with a panel of monoclonal antibodies. Twenty-nine were small cell lung carcinoma (SCLC), 15 adenocarcinomas, 17 squamous carcinomas and 10 large cell carcinomas. Three of the monoclonal antibodies recognize different cytokeratins, three recognize other epithelial antigens and one recognizes a neural antigen. Both formalin-fixed and cryopreserved tumours were studied using an indirect immunoperoxidase method. 23/29 SCLC reacted with all but one of the antibodies which recognize epithelial antigens. This staining was similar to that seen in non small cell lung carcinomas (NSCLC) and provides further evidence that SCLC are true epithelial tumours. All but one of the SCLC stained with the antibody recognizing a neural antigen. This antibody did not stain squamous or adenocarcinomas. However, four of the large cell carcinomas stained well with this antibody, suggesting that SCLC and some large cell carcinomas share a common pathway of differentiation. There were variations of staining seen both within and between tumours. This has obvious implications if immunotargetting with monoclonal antibodies is to be used diagnostically or therapeutically.     

miR-206及Bcl-2蛋白在非小细胞肺癌中的表达及意义

目的探讨mi R-206及Bcl-2蛋白在非小细胞肺癌(NSCLC)的表达,以及抑制或过表达mi R-206对Bcl-2蛋白表达的影响。方法实时定量PCR方法检测NSCLC组织mi R-206的表达水平,Western blot方法检测NSCLC组织Bcl-2蛋白表达;将mi R-206 inhibitors及mi R-206 mimics转染至A549细胞,通过CCK-8法测定A549细胞增殖能力的变化,Western blot方法检测转染后Bcl-2蛋白表达变化。结果 mi R-206在非小细胞肺癌组织中的表达量较癌旁组织明显降低(0.01),Bcl-2蛋白在癌组织中较癌旁组织表达明显增高(0.05);转染mi R-206 inhibitor的细胞增殖能力与对照组相比显著升高,细胞内Bcl-2蛋白水平亦明显升高(0.05),转染mi R-206 mimics的细胞增殖能力与对照组相比显著降低,细胞内Bcl-2蛋白水平亦明显降低(0.05)。结论mi R-206在NSCLC组织中低表达,mi R-206抑制能够促进A549细胞增殖和Bcl-2蛋白表达。     

miR-183及Akt1蛋白在非小细胞肺癌中的表达及其意义

目的探讨mi R-183及Akt1蛋白在非小细胞肺癌(NSCLC)组织中的表达,以及抑制或过表达mi R-183对Akt1蛋白表达的影响。方法实时定量PCR方法检测NSCLC组织mi R-183m RNA及Akt1m RNA的表达水平,Western blot方法检测NSCLC组织Akt1蛋白表达;将mi R-183 inhibitors及mi R-183 mimics转染至A549细胞,实时定量PCR方法及Western blot方法分别检测转染后Akt1 m RNA及蛋白的表达变化。结果 mi R-183m RNA在非小细胞肺癌组织中的表达量较癌旁组织明显降低(0.01),Akt1 m RNA及蛋白在癌组织中较癌旁组织表达明显增高(0.05);转染mi R-183 inhibitors的细胞内Akt1蛋白与m RNA水平与对照组相比显著升高(0.05),转染mi R-183 mimics的细胞内Akt1蛋白与m RNA水平对照组相比显著降低(0.05)。结论 mi R-183在NSCLC组织中低表达,并能够调节Akt1的表达。     

miR-23a/b在非小细胞肺癌的表达及临床意义

目的研究miR-23a和miR-23b在非小细胞肺癌中的表达特征,并探讨其临床意义。方法收集157例非小细胞肺癌手术切除标本及50例癌旁组织标本,采用Real time PCR方法检测miR-23a和miR-23b在非小细胞肺癌及其癌旁组织中的表达,分析二者表达的相关性,并探讨其表达与临床病理特征及其预后的关系。结果 miR-23a和miR-23b在非小细胞肺癌组织中的表达水平均高于癌旁肺组织,二者在非小细胞肺癌组织中表达呈正相关(r=0.351,P0.001)。miR-23a和miR-23b联合高表达与淋巴结有无转移(P0.001)、远处转移(P=0.001)及临床分期(P=0.002)相关,而与年龄、性别、组织类型及组织分化程度无显著相关性(P0.05)。KaplanMeier分析显示miR-23a和miR-23b联合高表达组患者生存期显著低于单独高表达组或联合低表达组(P=0.009),Cox风险比例模型分析显示miR-23a和miR-23b联合高表达为非小细胞肺癌患者的危险因素。结论 miR-23a和miR-23b在非小细胞肺癌中异常高表达可能是潜在的肺癌预后分子标志物。     

科罗索酸通过抑制Hippo-YAP信号转导通路促进非小细胞肺癌细胞的凋亡

目的：研究科罗索酸对肺癌细胞增殖与凋亡的影响及其机制。方法：MTT、Caspase3/7活性检测、Western blot等方法检测不同浓度的科罗索酸是否影响肺癌细胞株A549的增殖和凋亡;Western blot、免疫荧光实验等方法检测科罗索酸对肺癌细胞A549 Hippo通路中YAP蛋白进行定量和定位的检测。结果：MTT结果显示科罗索酸能够抑制肺癌细胞株A549的增殖,半抑制浓度为40 μmol/L;Caspase活性检测结果显示科罗索酸能够促进肺癌细胞株A549的凋亡,半致死浓度为40 μmol/L;Western blot、免疫荧光试验结果显示科罗索酸能明显抑制肺癌细胞株A549中YAP蛋白的表达。结论：科罗索酸可能通过调控Hippo-YAP信号通路抑制肺癌细胞的增殖并促进其凋亡。     

Human papillomavirus (HPV) and Merkel cell polyomavirus (MCPyV) in non small cell lung cancer

Certain types of human papillomavirus (HPV) induce cancers, especially cervical cancers in women. A meta-analysis of the literature suggests that HPV is also associated with 20%–25% of non small cell lung carcinoma (NSCLC). Merkel cell Polyomavirus (MCPyV) causes most Merkel cell carcinomas in immunocompromised hosts, and is associated with some squamous carcinomas of skin in immunocompetent individuals. Since both oncogenic viruses appear to involve the tonsils and, therefore, have clear access to the lungs, we examined that the possible association of HPV and MCPyV infections with lung cancers, especially, NSCLC. DNAs were extracted from 51 frozen tissues from 30 lung cancer patients, and examined for the presence of HPV and MCPyV by PCR and DNA sequencing analysis. Clinical data was correlated with the viral status. HPVs were only detected in 5 adenocarcinomas (16.7% of all lung cancers examined). Three were positive for HPV-16, 1 for HPV-11 and 1 had an unknown HPV type DNA. None was identified in benign tissue. MCPyV DNA was detected in 5 NSCLCs (16.7%). Three of the 5 were identified in squamous carcinomas, 1 in adenocarcinoma, and 1 in an unspecified NSCLC. Two additional samples were positive for MCPyV DNA within benign adjacent lung tissue only. In one adenocarcinoma, HPV-11 was identified in an adenocarcinoma, and MCPyV DNA was detected in the adjacent “benign” tissue. HPV and MCPyV were directly associated with 33.3% of NSCLC. Further studies are necessary to determine if polyomavirus and papillomavirus are necessary risk factors for some cases of NSCLC.