野生型P53、P16基因协同对肺腺癌细胞系生长抑制作用的研究

为了探讨野生型P53基因及P16基因在恶性肿瘤基因治疗中的作用,用腺病毒为载体将野生型P53基因转入高、低转移的肺腺癌细胞系Anip973、AGZY83-a和经野生型P16基因质粒转染的高、低转移肺腺癌细胞系Anip973（Anip973P16）、AGZY83-a（AGZY83-aP16）。对各组转染细胞进行生长曲线、MTT生长抑制率、原位末端标记、Western-blotting等技术检测分析。结果发现（1）野生型P53蛋白的过表达对上述肺腺癌细胞系均呈现出较强的生长抑制作用。（2）野生型P53蛋白的过表达对高转移肺癌细胞系Anip973的抑制作用明显高于低转移细胞系AGZY83-a。（3）野生型p53蛋白的过表达对经野生型P16基因转染的高、低转移的肺癌细胞Anip973、AGZY83-a抑制作用明显高于未经P16基因转染的细胞。野生型P53基因可以作为肺腺癌基因治疗的候选基因。肿瘤抑制基因P53、P16的联合转染可能是对肺腺癌进行基因治疗的有效手段。 Abstract：To investigate the suppression effect of tumor suppressor genes in lung adenocarcinoma cell lines,we transferred a pair of lung adenocarcinoma cell lines with different metastasis potential,Anip973(High-metastasis potential cell line) and AGZY83-a (Low-metastasis potential cell line)and this pair of cell lines transfected with P16 gene:AGZY83-a P16 and Anip973 P16 with wild type P53 gene with adenovirus vector.The suppression effects of P53 gene were evaluated by cell growth curve,MTT,western-blotting analysis and TUNEL technique.Overexpression of wild-type P53 gene in AGZY83-a,Anip973,Anip973 P16 and AGZY83-a P16 inhibited the growth of these four kinds of lung cancer cells and induced apoptosis of the cells.The suppression effect of P53 gene in Anip973 and Anip973 P16 was higher than AGZY83-a and AGZY83-a P16 while co-expression of P53 and　P16 in this pair of cell lines inhibited the cells more efficiently comparing with the expression of P53 alone.Wild-type P53 gene might act as a candidate gene in lung adenocarcinoma gene therapy while co-transfection of P53 and P16 genes was a more effective method.     

一个与非小细胞肺癌转移相关的基因――RAB5A基因

采用mRNA差异展示技术（mRNA DD）研究具有相同细胞来源,但转移能力高低不同的人肺腺癌细胞系AGZY83-a（低转移）和Anip973(高转移),分析在两个细胞系中基因差异表达的情况,发现在高转移细胞系中有RAB5A基因的表达。该基因为蛋白质入胞信号的调控者,为RAS超家族成员。为进一步证实其转录表达的调控改变情况,以及RAB5A高表达的临床意义,进一步采用RT-PCR和免疫组织化学的方法检测了50例临床非小细胞肺癌的手术标本,结果表明,RAB5A的表达有随转移发生而增强的趋势,而RAB5A的蛋白表达程度在有转移的病例中明显增强(P<0.05)。 Abstract：　Using mRNA differential display (mRNA DD)techniques, we analyzed the differences of gene expression between two human lung adenocarcinoma cell lines,AGZY83-a and Anip973. Anip973 was isolated from AGZY83-a, but manifested much higher metastatic potential than the parent line. The results showed that there were significant differences on gene expression between the two cell lines and that there was over-expression of RAB5Agene in the Anip973 cell line. The product of RAB5Agene was recognized as signal regulators of endocytotic pathway and protein trafficking at the cell surface, and belong to a member of the RAS superfamily. Furthemore, we compared to the expression of RAB5Agene and RAB5Aprotein in clinical samples of 50 cases non-small lung carcinoma and nearby lymph node by means of RT-PCR and immunohistochemistry method. The results indicated that the high expression of RAB5Ain metastatic tumor and the enhancement level of RAB5Ain metastatic tumor and the enhancement level of expression were corresponded with the increase of metastatic degree. And there were significance of difference on the expression degree of RAB5Aprotein between non-small lung carcinoma with metastasis and non- metastasis (P<0.05).     

Upregulation of BNIP3 promotes apoptosis of lung cancer cells that were induced by p53

It has been shown that p53 induces cell apoptosis and the Bcl-2 family plays key roles in this process. However, the molecular mechanism of p53 apoptotic pathway is still unclear. Here, we show that overexpression of exogenous wild-type p53 induced apoptosis in lung cancer cells and high metastasis potential cells had a faster rate of apoptosis than low metastasis potential cells. The expression of pro-apoptotic gene BNIP3 was increased significantly both in Anip973 and 95D cell lines which have high metastasis ability, but not AGZY83-a or little increased in 95C cell lines which possess low metastasis ability. Overexpression of BNIP3 increases apoptotic rate induced by p53 in AGZY83-a cells. Blocking the expression of BNIP3 by siRNA in Anip973 cells decreased apoptotic rate mediated by p53. Taken together, these data suggest that high level expression of BNIP3 mediated rapid apoptosis that was triggered by p53 in lung cancer cells.     

Id 基因在多种肺癌细胞中的表达及意义

目的：研究Id基因在肺癌和永生化支气管上皮细胞中的表达,探讨其在肺癌细胞中表达的意义。方法：利用半定量RT—PCR和Western blot方法检测多种肺癌和永生化支气管上皮细胞中Id1—Id4 mRNA和Id1—Id4蛋白的表达。结果：A549、NCI—H460、NCI—H446、SK—MES—1、Anip973中Id1-Id3 mRNA均高表达,Id1相对表达较强;而AGZY和MP-184中未表达Id1-Id3 mRNA;腺癌细胞均表达了Id4 mRNA,而NCI-H446、SK—MES—1未表达Id4mRNA。A549,NCA—H460,NCA—H446,SK—MES—1,Anip973中Id1,Id2,Id3蛋白均高表达,A549,NCA—H446,Anip973中Id2的表达高于NCA—H460,SK—MES—1;A549,NCA—H460,Anip973有出的高表达,NCI—H446,SK—MES—1无Id4的表达,Id1-Id4在AGZY和MP-184中均耒表达。结论：4种Id基因均作为癌基因在肺癌的发生发展中发挥作用,Id1,Id2,Id3与肺癌细胞的恶性程度以及增殖和转移密切相关,Id4可做为肺腺癌的检测标志物。     

13q14断裂重排与非小细胞肺癌转移潜能关系的研究

肿瘤转移的细胞经常存在染色体数目异常和结构畸变,在多种有转移潜能的肿瘤细胞中都涉及到13q14的异常。以往研究表明在同一组织来源但转移潜能不同的肺腺癌细胞系AGZY83-a和Anip973中存在13q14的断裂重排。采用mRNA差异展示技术（mRNA DD）分析这一对细胞系得到的差异表达基因BRI基因位于13q14。为了进一步分析肺癌细胞的转移潜能与13q14断裂重排间的关系,采用13q涂染探针对具有不同转移潜能的非小细胞肺癌细胞系PAa、SPC-1-A和95D中期分裂相进行G显带后的荧光原位杂交分析。结果发现在3个肺癌细胞系中有多种13号染色体长臂的结构异常,其中此3个细胞系均涉及13q32-33的频发断裂。但是低转移肺癌细胞系PAa、SPC-1-A均未涉及13q14的断裂,而高转移肺癌细胞系95D的两种细胞克隆均可见13q14的断裂。提示13q14断裂点与肺癌细胞的转移能力有一定的相关性,两者之间的遗传学意义需要进一步研究探索。     

rab5a基因在肿瘤转移中的作用研究

为研究rab5a基因在肿瘤转移机制中的作用,将该基因稳定转染至低转移肺腺癌细胞系AGZY83-a中,采用Superarray肿瘤转移相关基因微芯片分析rab5a对肿瘤转移相关基因的表达影响,共获得了5个差异表达基因,rab5a基因促进s100a4的表达,同时抑制了nm23a、rac1、cst3、col4a2等基因的表达,并分别在RNA及蛋白水平进行验证,确认rab5a基因影响了肿瘤转移的多个途径,促进了肿瘤细胞转移能力增强。     

Decrease of PDSS2 expression,a novel tumor suppressor,in non-small cell lung cancer

Background: Prenyl diphosphate synthase subunit 2 (PDSS2) gene has recently been reported as a potential tumor suppressor. The association of PDSS2 and non-small cell lung cancer (NSCLC) has not been known. Methods: To investigate its association with NSCLC, we examined the expression level of PDSS2 in 28 paired clinical samples of non-small cell lung cancer tissues and surrounding normal tissues. Results: PDSS2 was constitutionally expressed in normal lung tissues regardless of sex, race and smoking history. An overall decreased PDSS2 expression was found in the tumor tissues compared to surrounding normal tissues. Decrease in PDSS2 expression was more severe in poorly and poor-to-moderately differentiated lung cancers, while the decrease was not significant in moderately to well-differentiated tumors. Moreover, the expression of PDSS2 decreased more in higher pathological stage, and in patients with lymph node metastasis. The decrease in PDSS2 expression in tumor tissues was not related to sex or histological type of NSCLC, but was related to smoking history. No correlation has been found between PDSS2 and the clinical factors of EGRF, Ki-67 and p53. Conclusion: Taken together, decreased expression of PDSS2 in NSCLC was evident. This is an initial report for the expression of PDSS2 in relation to different factors in lung cancer. Loss of PDSS2 could serve as a potential biomarker in NSCLC development. The role of PDSS2 as a tumor suppressor, and the mechanism of its potential anti-tumor action in NSCLC warrant further investigation.     

水飞蓟宾诱导肺腺癌Anip973 细胞凋亡的分子机制研究

目的:探讨水飞蓟宾诱导肺腺癌Anip973细胞系细胞凋亡的分子机制。方法:采用MTT法、倒置显微镜和电子显微镜等形态学检测以及流式细胞仪(FCM)技术检测、DNALadder分析、凋亡分子PARP的表达检测细胞凋亡,同时进行凋亡相关蛋白Bax、Bcl-2、caspase-3和caspase-9表达活性分析。结果:(1)水飞蓟宾对人肺腺癌Anip973细胞系细胞的增殖有显著抑制作用;(2)水飞蓟宾作用Anip973细胞48h后,随着浓度的增加,倒置显微镜下可见细胞数目减少,胞体变小、变圆,到高浓度时出现较多的死亡细胞;(3)扫描电镜观察发现,随着水飞蓟宾作用浓度的增加,Anip973细胞中出现增多的凋亡细胞,凋亡细胞表现出典型的超微结构特征;(4)流式细胞仪检测的结果发现,随着药物作用时间的延长,Anip973细胞的G1期细胞比例增多,S期细胞明显减少,G2期细胞略有减少,并出现明显的凋亡峰。(5)水飞蓟宾作用后的Anip973细胞出现明显的DNALadder和PARP降解增加等凋亡特征;(6)水飞蓟宾作用后,Anip973细胞中的凋亡相关蛋白Bax表达增加、caspase-3和caspase-9酶活性增加,而Bcl-2表达降低。结论:水飞蓟宾在体外有抑制人肺腺癌细胞Anip973的增殖作用,并通过激活线粒体依赖的caspase凋亡通路,诱导其凋亡。     

水飞蓟宾诱导肺腺癌Anip973细胞凋亡的分子机制研究

目的：探讨水飞蓟宾诱导肺腺癌Anip973细胞系细胞凋亡的分子机制。方法：采用MTT法、倒置显微镜和电子显微镜等形态学检测以及流式细胞仪（FCM）技术检测、DNALadder分析、凋亡分子PARP的表达检测细胞凋亡,同时进行凋亡相关蛋白Bax、Bcl-2、caspase-3和caspase-9表达活性分析。结果：（1）水飞蓟宾对人肺腺癌Anip973细胞系细胞的增殖有显著抑制作用;（2）水飞蓟宾作用Anip973细胞48h后,随着浓度的增加,倒置显微镜下可见细胞数目减少,胞体变小、变圆,到高浓度时出现较多的死亡细胞;（3）扫描电镜观察发现,随着水飞蓟宾作用浓度的增加,Anip973细胞中出现增多的凋亡细胞,凋亡细胞表现出典型的超微结构特征;（4）流式细胞仪检测的结果发现,随着药物作用时间的延长,Anip973细胞的G1期细胞比例增多,S期细胞明显减少,G2期细胞略有减少,并出现明显的凋亡峰。（5）水飞蓟宾作用后的Anip973细胞出现明显的DNALadder和PARP降解增加等凋亡特征;（6）水飞蓟宾作用后,Anip973细胞中的凋亡相关蛋白Bax表达增加、caspase-3和caspase-9酶活性增加,而Bcl-2表达降低。结论：水飞蓟宾在体外有抑制人肺腺癌细胞Anip973的增殖作用,并通过激活线粒体依赖的caspase凋亡通路,诱导其凋亡。     

miR-204通过抑制OGT的表达调控非小细胞肺癌细胞的增殖和迁移

目的:探究miR-204和O-连接N-乙酰氨基葡萄糖转移酶(O link N-acetylglucosamine transferase,OGT)对非小细胞肺癌(Non-small cell lung cancer,NSCLC)细胞增殖和转移的影响,并深入分析其可能机制。方法:采用Oncomine及KM-Ploter数据库分析OGT在肺癌组织中的表达及与肺癌患者预后的关系;采用慢病毒转染人非小细胞肺癌A549及永生化人肺支气管上皮细胞BEAS-2B,分别构建OGT稳定下调和过表达的细胞系,利用CCK-8、平板克隆和裸鼠皮下成瘤实验检测细胞增殖的情况,划痕实验、Transwell实验和裸鼠尾静脉注射肺转移模型检测细胞转移的情况。利用数据库分析可能参与OGT调控的microRNA,并用双荧光素酶报告基因验证。利用TCGA数据库分析miR-204在肺癌中的表达情况,并在30例肺癌组织及其对应癌旁组织中分析miR-204与OGT之间的相关性。结果:OGT在肺癌组织中呈高表达,且与患者的不良预后相关(HR=1.22,P 0.01);OGT的表达上调肺癌细胞的增殖和转移;miR-204可以负向调控OGT的表达,且miR-204在肺癌组织中表达水平显著低于癌旁组织,在肺癌组织中miR-204的水平与OGT的表达水平呈负相关(R~2=-0.4729,P 0.01)。结论:在非小细胞肺癌中,miR-204的降低通过上调OGT的表达促进肺癌的增殖和转移。     

lncRNA DGCR5在非小细胞肺癌组织中的表达及其与临床病理特征的相关性分析

摘要 目的：探究lncRNA DGCR5在非小细胞肺癌(NSCLC)组织中的表达及其与临床病理特征的相关性。方法：选取2020年1月至2021年12月在我院肿瘤科收治的进行手术治疗的NSCLC患者86例,在手术期间从患者获得肿瘤和非肿瘤的肺癌旁组织样本。采用qRT-PCR测定肿瘤组织及癌旁组织中lncRNA DGCR5表达水平。分析lncRNA DGCR5表达水平与NSCLC患者性别、年龄、临床分期、T分期、N分期等临床病理参数的关系,lncRNA DGCR5表达水平与患者预后总生存期（OS）和无进展生存期（PFS）的关系。结果：与癌旁组织相比,lncRNA DGCR5在NSCLC肿瘤组织中的表达水平相对较低,差异具有统计学意义（P<0.01）。lncRNA DGCR5表达与肿瘤分化程度、TNM分期、肿瘤体积、淋巴转移和远处转移之间存在明显相关性,差异具有统计学意义（P<0.05）。采用Kaplan-Meier法进行生存分析,研究发现lncRNA DGCR5高表达组中位OS及中位DFS分别显著高于lncRNA DGCR5低表达组（P<0.05）。低分化程度、II+ IIIa临床分期、N1-N3淋巴转移、远处转移、及lncRNA DGCR5 低表达均与NSCLC患者总生存率和无进展生存率相关。结论：LncRNA DGCR5在NSCLC患者肿瘤组织中的表达量降低,NSCLC患者血LncRNA DGCR5表达水平与分化程度、TNM分期、淋巴转移、远处转移及预后具有相关性。LncRNA DGCR5可作为早期诊断和治疗NSCLC的新型生物标志物。     

Exosomal miR-1260b derived from non-small cell lung cancer promotes tumor metastasis through the inhibition of HIPK2

Tumor-derived exosomes (TEXs) contain enriched miRNAs, and exosomal miRNAs can affect tumor growth, including cell proliferation, metastasis, and drug resistance through cell-to-cell communication. We investigated the role of exosomal miR-1260b derived from non-small cell lung cancer (NSCLC) in tumor progression. Exosomal miR-1260b induced angiogenesis by targeting homeodomain-interacting protein kinase-2 (HIPK2) in human umbilical vein endothelial cells (HUVECs). Furthermore, exosomal miR-1260b or suppression of HIPK2 led to enhanced cellular mobility and cisplatin resistance in NSCLC cells. In patients with NSCLC, the level of HIPK2 was significantly lower in tumor tissues than in normal lung tissues, while that of miR-1260b was higher in tumor tissues. HIPK2 and miR-1260b expression showed an inverse correlation, and this correlation was strong in distant metastasis. Finally, the expression level of exosomal miR-1260b in plasma was higher in patients with NSCLC than in healthy individuals, and higher levels of exosomal miR-1260b were associated with high-grade disease, metastasis, and poor survival. In conclusion, exosomal miR-1260b can promote angiogenesis in HUVECs and metastasis of NSCLC by regulating HIPK2 and may serve as a prognostic marker for lung cancers.Subject terms: Non-small-cell lung cancer, Metastasis, miRNAs     

非小细胞肺癌组织PRRX1、VASH-1与微血管密度、临床病理参数和预后的关系研究

摘要 目的：探讨非小细胞肺癌（NSCLC）组织配对相关同源框蛋白1（PRRX1）、血管抑制蛋白1（VASH-1）与微血管密度（MVD）、临床病理参数和预后的关系。方法：选择2018年1月至2020年1月辽宁省金秋医院行手术切除的156例NSCLC患者的癌组织及癌旁正常组织标本。应用免疫组织化学染色法检测癌组织及癌旁组织PRRX1、VASH-1的阳性表达率,并进行MVD计数。比较PRRX1阳性表达组/阴性表达组、VASH-1阳性表达组/阴性表达组MVD计数。分析PRRX1、VASH-1与NSCLC患者病理参数的关系。随访3年,应用Kaplan-Meier生存曲线分析PRRX1、VASH-1阳性/阴性表达与NSCLC患者预后的关系。结果：与癌旁组织相比,NSCLC患者癌组织PRRX1阳性表达率降低,VASH-1阳性表达率升高（P<0.05）。与PRRX1阴性NSCLC患者相比,PRRX1阳性NSCLC患者癌组织MVD降低,与VASH-1阴性NSCLC患者相比,VASH-1阳性NSCLC患者癌组织MVD升高（P<0.05）。与TNM I~II期、无淋巴结转移NSCLC患者的癌组织相比,TNM Ⅲ A期、淋巴结转移NSCLC患者的癌组织中PRRX1阳性表达率降低,VASH-1阳性表达率升高（P<0.05）。Kaplan-Meier法分析显示,PRRX1阳性组3年总体生存率（OS）、3年无病生存率（DFS）高于PRRX1阴性组（P<0.05）,VASH-1阴性组3年OS、3年DFS高于VASH-1阳性组（P<0.05）。结论：NSCLC患者的癌组织中PRRX1阳性表达率降低,VASH-1阳性表达率升高,与淋巴结转移、TNM分期及不良预后有关。     

TRIM28, a new molecular marker predicting metastasis and survival in early-stage non-small cell lung cancer

TRIM28 is a universal corepressor for Kruppel-associated box zinc finger proteins. In this study, we demonstrated the expression of TRIM28 gene was significantly higher in cancerous tissues than in noncancerous tissues (P < 0.001). TRIM28 knockdown resulted in a decrease in cell proliferation in liquid media as well as in soft agar. The proliferation rate was impaired and the cell cycle progression was inhibited after knockdown of TRIM28 in non-small cell lung cancer cell lines PAa and SK-MES-1. We used real-time polymerase chain reaction to detect circulating cancer cells in 138 non-small cell lung cancer patients. The overall positive detection rate was 30.4% (42 of 138) in peripheral blood of NSCLC patients and was 29.9% (29 of 97) in early-stage patients. In a 70-month follow-up study, 20 of 29 patients (69.0%) in TRIM28 positive group had recurrence and/or metastasis, significantly higher (P = 0.004) than in the TRIM28 negative group (25 of 68, 36.8%). In addition, non-small cell lung cancer patients whose circulating cancer cells expressed TRIM28 suffered shorter tumor-specific survival compared with those with absent TRIM28 expression (P < 0.001). Results of our study showed that TRIM28 provides a survival advantage to lung cancer cells and may be a new marker to predict metastasis and prognosis in early-stage non-small cell lung cancer patients.     

埃兹蛋白的表达与非小细胞肺癌转移的关系研究

目的:探究埃兹蛋白(Ezrin)的表达与非小细胞肺癌转移的关系。方法:通过免疫组化检测Ezrin在有无转移的非小细胞肺癌组织中的表达差异,通过细胞免疫组化、western-blot、RT-PCR检测Ezrin在不同转移潜能肺癌细胞系中的表达差异,通过transwell考察Ezrin对不同转移潜能癌细胞侵袭和迁移能力的影响。结果:Ezrin蛋白在有转移的非小细胞肺癌组织中的表达水平明显高于无转移的肺癌组织,在高转移潜能肺癌细胞系中的表达高于低转移潜能细胞系,受抑制时会削弱高转移潜能肺癌细胞的迁移和侵袭能力,过表达时会增强低转移潜能肺癌细胞的侵袭和迁移能力。结论:Ezrin可能在非小细胞肺癌及其转移中发挥重要作用。     

SBF2-AS1: An oncogenic lncRNA in small-cell lung cancer

The long noncoding RNAs (lncRNAs) SBF2 antisense RNA 1 (SBF2-AS1) was found to act as an oncogenic lncRNA in non–small-cell lung cancer (NSCLC), but the role of SBF2-AS1 in small-cell lung cancer (SCLC) was still unclear. The purpose of this study was to provide the clinical significance and biological function of SBF2-AS1 in SCLC. In our results, SBF2-AS1 was found to be upregulated in SCLC tissues compared with NSCLC tissues or adjacent normal lung tissues. Besides, SBF2-AS1 expression was also elevated in SCLC cell lines compared with the normal bronchial epithelial cell line or NSCLC lines. Moreover, high expression of SBF2-AS1 was associated with clinical stage, tumor size, lymph node metastasis and distant metastasis in SCLC patients. Survival analysis showed SCLC patients with high expression of SBF2-AS1 had shorter overall survival than patients with low expression of SBF2-AS1, and high expression of SBF2-AS1 acted as an independent poor prognostic factor for overall survival in SCLC patients. The study in vitro suggested inhibition of SBF2-AS1 obviously depressed cell proliferation, migration, and invasion in SCLC. In conclusion, SBF2-AS1 acts as a novel oncogenic lncRNA in SCLC.     

Knockdown of LncRNAZFAS1 suppresses cell proliferation and metastasis in non-small cell lung cancer

ABSTRACT

To evaluate the effects of LncRNAZFAS1 on cell proliferation and tumor metastasis in non-small cell lung cancer (NSCLC), we detected the expression level of LncRNAZFAS1 in NSCLC-related tissues and cells. qRT-PCR results revealed that LncRNAZFAS1 in tumor tissues was significantly higher than that in normal lung tissue, especially significantly up-regulated in stage III / IV and in metastatic NSCLC tissues. LncRNAZFAS1 expression was dramatically up-regulated in 4 NSCLC-related cells (A549, SPC-A1, SK-MES-1, and NCI-H1299), with having the highest expression level in A549 cells. Furthermore, we implemented a knockdown of LncRNAZFAS1 in A549 cells, and the results of CCK8 and Transwell assays suggested that knockdown of LncRNAZFAS1 significantly inhibited NSCLC cell proliferation and metastasis. Next, we constructed a tumor xenograft model to evaluate the effect of LncRNAZFAS1 on the NSCLC cell proliferation in vivo. The results indicated that knockdown of LncRNAZFAS1 dramatically inhibited A549 cells proliferation and repressed tumor growth. Additionally, knockdown of LncRNAZFAS1 drastically weakened the expressions of MMP2, MMP9 and Bcl-2 proteins, whereas noticeably strengthened the expression of BAX protein. Our results altogether suggest that knockdown of LncRNAZFAS1 has a negative effect on the proliferation and metastasis of NSCLC cell, which implying LncRNAZFAS1 is a potential unfavorable biomarker in patients with NSCLC.     

Expression of THOP1 and Its Relationship to Prognosis in Non-Small Cell Lung Cancer

Background

The study was designed to detect the expression level of thimet oligopeptidase (THOP1) protein in non-small cell lung cancer (NSCLC) and investigate its correlation with clinicopathologic features and prognosis.

Methods

Immunohistochemical staining was used to determine the expression of THOP1 protein in 120 NSCLC specimens and 53 distant normal lung tissues. Quantitative real-time PCR and western blotting were employed to measure the expression of THOP1 in 16 pairs of primary NSCLC and corresponding normal tissues.

Results

Analysis of immunohistochemical staining suggested low THOP1 expression was found in 71 (59.2%) of the 120 NSCLC specimens and significantly correlated with positive lymph node metastasis (P = 0.048). However, low THOP1 expression was found in 22 (41.5%) of the 53 normal lung tissues. Chi-square test suggested that the expression of THOP1 was significantly higher in the normal lung tissues than that in the NSCLC specimens (P = 0.032). Real-Time PCR and western blotting showed that NSCLC specimens had decreased THOP1 mRNA and protein expression compared to corresponding normal tissues. Univariate analysis demonstrated that low THOP1 expression significantly predicted decreased 5-year disease-free survival (P = 0.038) and overall survival (P = 0.017). In addition, positive lymph node metastasis (P = 0.025) and advanced TNM stage (P = 0.009) significantly predicted decreased 5-year overall survival. However, multivariate Cox regression analysis showed that only low THOP1 expression retained its significance as an independent prognostic factor for unfavorable 5-year disease-free survival (P = 0.046) and overall survival (P = 0.021).

Conclusions

THOP1 may have clinical potentials to be employed as a promising biomarker to identify individuals with better prognosis and a novel antitumor agent for therapy of patients with NSCLC.