Immunohistochemical analysis and comparison of napsin A,TTF1, SPA and CK7 expression in primary lung adenocarcinoma

We investigated the expression of napsin A, TTF1, SPA and CK7 in primary lung adenocarcinoma (ADC) and compared antibody specificity and sensitivity. We used immunohistochemistry (IHC) using antibodies directed against napsin A, TTF1, SPA, and CK7 on cancer tissue from 454 patients diagnosed with lung cancer. In primary lung ADC, napsin A exhibited a sensitivity of 87.25% and specificity of 95.02%, TTF1 exhibited a sensitivity of 80.72% and specificity of 90.48%, SPA exhibited a sensitivity of 60.46% and specificity of 94.87% and CK7 exhibited a sensitivity of 100% and specificity of 80.31%. The order of sensitivity of the four indicators was CK7 > napsin A > TTF1 > SPA, and the order of specificity was napsin A > SPA > TTF1 > CK7. We can select effective indicators for diagnostic and differential diagnostic purposes based on differences in sensitivity and specificity.     

子宫内膜癌C-erbB-2和TTF-1的表达及其与预后的关系

目的:探讨子宫内膜癌组织中C-erbB-2和TTF-1的表达及其与患者临床预后的关系。方法:应用免疫组化S-P法检测正常子宫内膜组织、子宫内膜不典型增生组织、子宫内膜癌组织中C-erbB-2与TTF-1的表达,并分析二者与子宫内膜癌临床病理特征及患者临床预后的关系。结果:子宫内膜癌组织、子宫内膜不典型增生组织、正常子宫内膜组织中C-erbB-2的阳性表达率分别为56.65%、36.67%、16.67%(P0.05),TTF-1的的阳性表达率分别为33.75%、53.33%、70.00%(P0.05)。C-erbB-2的阳性表达与子宫内膜癌的临床病理分期、组织分化、肌层浸润及有无淋巴结转移呈显著正相关(P0.05),与其病理类型无显著相关性(P0.05)。TTF-1的阳性表达与子宫内膜癌的病理分期、组织分化、肌层浸润深度负相关(P0.05),但与有无淋巴结转移及病理类型均无显著相关性(P0.05)。C-erbB-2和TTF-1在子宫内膜癌组织中表达无明显相关性(P=0.303)。子宫内膜癌中C-erbB-2阳性表达者五年生存率(80.90%)显著低于C-erbB-2阴性表达者(93.80%)(P0.05),TTF-1阳性表达者五年生存率(87.70%)略高于TTF-1阴性表达者(85.70%),但差异无统计学意义(P0.05)。结论:子宫内膜癌组织中C-erbB-2的表达显著上调,而TTF-1的表达明显下调,二者表达异常均参与了子宫内膜癌的发生和发展,且C-erbB-2的表达上调与子宫内膜癌患者的不良预后相关。     

MSK2 promotes proliferation and tumor formation in squamous cervical cancer via PAX8/RB-E2F1/cyclin A2 axis

Patients with cervical cancer have abnormal cell proliferation and invasion after many years of latency. However, the precise mechanisms remain unclear. Mitogen- and stress-activated kinase 2 (MSK2) is a serine/threonine kinase which displays a phenotype that promotes tumor growth and metastasis in many different types of tumors. The aim of the present study was to determine the effects of MSK2 on the proliferation of cervical cancer cells and elucidate the signaling pathways through which MSK2 exerts its effects in the pathogenesis of squamous cell carcinoma (SCC). Our results confirmed that MSK2 expression was significantly upregulated in cervical cancer cells both in vivo and in vitro. We further found that the expression patterns of paired-box gene 8 (PAX8) and MSK2 were positively correlated in cervical cancer specimens. Moreover, MSK2 knockdown inhibited the phosphorylation of PAX8 and retinoblastoma protein (RB), and suppressed the sequential expressions of cell proliferation factors E2F1 and cyclin A2, resulting in the inhibition of SCC cell proliferation and tumor formation. Thus, this study demonstrates that MSK2 has oncogenic effects in the formation and development of SCC via the PAX8/RB-E2F1/cyclin A2 axis.     

The genomic alterations of lung adenocarcinoma and lung squamous cell carcinoma can explain the differences of their overall survival rates

In the US, lung carcinoma accounted for over 150,000 deaths in 2018 and the advances in increasing survival rates are still limited. In this study, we investigated the cohorts with lung adenocarcinoma (LUAD) or lung squamous cell carcinoma (LUSC) from The Cancer Genome Atlas to figure out the risk factors and genomic alterations that affected their prognosis. The histoclinical factors that differed between LUAD and LUSC were identified and the risk factors affecting the overall survival were figured out for both LUAD and LUSC. Next, the patterns of nucleotides substitutions and the mutational signatures were extracted to illustrate whether different mutational processes performed for them. Finally, the genes that had different frequencies of mutation were identified. LUAD and LUSC presented differences in histoclinical factors including age at the time of diagnosis, sex, smoking history, pathological T classification, and overall survival. This was caused by the distinct genomic alterations including the transition-to-transversion ratios, mutational signatures, and the frequently mutated genes. We proposed that the mutational signature associated with aging could be used to predict the prognosis of patients with LUAD. On the other hand, the AID/APOBEC family was associated with the prognosis of LUSC. Finally, SNTG1 and LRRK2 might be important in LUAD and LUSC, respectively.     

Transforming growth factor beta-1 enhances cytotoxic effect of doxorubicin in human lung adenocarcinoma cells of A549 line

Transforming growth factor beta-1 (TGFbeta-1) is a regulator of cell proliferation, differentiation and apoptosis. Doxorubicin (adriamycin), an anthracycline drug causing double-strand DNA breaks, is widely used in anticancer chemotherapy. Here we demonstrated that TGFbeta-1 enhanced cytotoxic (proapoptotic) action of doxorubicin towards cultured human lung carcinoma A549 cells. Western-blot analysis and immunocytochemistry were used to show that doxorubicin induced PARP degradation in A549 cells, and TGFbeta-1 enhanced that action of the drug. The obtained results suggest a possibility of biomodulating effect of TGFbeta-1 on tumor cell treatment with doxorubicin.     

Exploring and comparing of the gene expression and methylation differences between lung adenocarcinoma and squamous cell carcinoma

Lung cancer is one of the most frequently diagnosed malignant tumors and the main reason for cancer-related death around the world, whereas nonsmall cell lung cancer that consists two subtypes: lung adenocarcinoma (LUAD) and squamous cell carcinoma (LUSC) is responsible for an estimated 85% of all lung cancers. The current study aimed to explore gene expression and methylation differences between LUAD and LUSC. EdgeR was used to identify differentially regulated genes between normal and cancer in the LUAD and LUSC extracted from The Cancer Genome Atlas (TCGA), respectively, whereas SAM was used to find genes with differential methylation between normal and cancer in the LUAD and LUSC, respectively. Finally, Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analysis was performed to analyze the function which these genes enriched in. A total of 391 genes with opposite methylation patterns in LUAD and LUSC and four functional pathways were obtained (false discovery rate (FDR) < 0.1). These pathways mainly included fat digestion and absorption, phenylalanine metabolism, bile secretion, and so on, which were related to the airframe nutrition metabolic pathway. Moreover, two genes CTSE (cathepsin E) and solute carrier family 5 member 7 (SLC5A7) were also found, among which CTSE was overexpressed and hypomethylated in LUAD corresponding to normal lung tissues, whereas SLC5A7 showed the opposite in LUAD. In conclusion, this study investigated the differences between the gene expression and methylation patterns in LUAD and LUSC, and explored their different biological characteristics. Further understanding of these differences may promote the discovery and development of new, accurate strategies for the prevention, diagnosis, and treatment of lung cancer.     

Methylation of CLEC14A is associated with its expression and lung adenocarcinoma progression

Our main objective is probing the effect of methylation of CLEC14A on its expression and lung adenocarcinoma (LUAD) progression. Microarray analysis was utilized to screen out differentially downregulated genes with hypermethylation in LUAD tissues. The CLEC14A expression level was measured by western blot analysis and qRT-PCR. Methylation-specific-PCR was performed to evaluate methylation status of CLEC14A. The 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromid (MTT) assay was used to check the relation between CLEC14A expression and cell proliferation. Cell cycle, cell apoptosis, migration, and invasion were respectively detected by the flow cytometry assay, wound healing assay, and transwell assay. Tumor xenograft models were established for investigating the effect of CLEC14A on tumor formation. CLEC14A expression in LUAD tissues was impaired compared with that in adjacent tissues, and CLEC14A promoter was highly methylated in LUAD. Overexpressing CLEC14A or inhibiting the methylation level of CLEC14A in A549 and LTEP-a-2 cells impeded the duplication of LUAD cells, promoted apoptosis, attenuated cell migration, and invasion ability, and arrested cell cycle at the G0/G1 phase. Overexpression of CLEC14A inhibited tumorigenesis of LUAD cells in nude mice. The promoter of CLEC14A is methylated in LUAD, leading to downregulation of CLEC14A in LUAD. CLEC14A acts as an antitumor role in LUAD by suppressing cell proliferation, migration, invasion, promoting cell apoptosis, and reducing tumorigenicity in nude mice. Thus, the inhibition of CLEC14A methylation is a novel strategy for the clinic treatment of LUAD.     

TRB3 interacts with ERK and JNK and contributes to the proliferation,apoptosis, and migration of lung adenocarcinoma cells

Tribbles homolog 3 (TRB3) has been accounted for regulation of a few cell processes through interaction with other significant proteins. The molecular mechanisms underlying TRB3 in tumorigenesis in lung adenocarcinoma have not been entirely elucidated. The present study is aimed at determining the function and fundamental mechanisms of TRB3 in lung adenocarcinoma progression. TRB3 was highly expressed in A549 and H1299 cells and lung adenocarcinoma tissues compared with human bronchial epithelial cells (HBEpC) and adjacent normal lung tissues. Hypoxia significantly upregulated the expression of TRB3 protein in A549 and H1299 cells in a time-dependent way. Gene expression profiling interactive analysis data analysis indicated that patients with lung adenocarcinoma with excessive expression of TRB3 mRNA had fundamentally shorter survival time. TRB3 knockdown in A549 cells can inhibit cell proliferation and migration, and promote cell apoptosis. TRB3 knockdown reduced the expression of p-ERK and p-JNK, but did not affect the expression of p-P38 MAPK. TRB3 overexpression enhances the malignant transformation abilities of HBEpC such as cell proliferation, migration and colony formation, which could be reversed by U0126 and SP600125. TRB3 overexpression promotes the phosphorylation of extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) but was not affected by U0126 and SP600125. The results of coimmunoprecipitation experiments indicated that TRB3 binds directly to ERK and JNK. This study suggests that TRB3 has a potentially carcinogenic role in lung adenocarcinoma by binding to ERK and JNK and promoting the phosphorylation of ERK and JNK. TRB3 can be a possible therapeutic focus for lung adenocarcinoma.     

Serum TNFRII: A promising biomarker for predicting the risk of subcentimetre lung adenocarcinoma

Early diagnosis of lung adenocarcinoma requires effective risk predictors. TNFRII was reported to be related to tumorigenesis, but remained unclear in lung cancer. This research set out to investigate the relationship between the sTNFRII (serum TNFRII) level and the risk of lung adenocarcinoma less than 1 cm in diameter. Seventy-one pairs of subcentimetre lung adenocarcinoma patients and healthy controls were analysed through multiplex bead-based Luminex assay and found a significantly lower expression of sTNFRII in patients with subcentimetre lung adenocarcinoma than that in the healthy controls (P < .001), which was further verified through ONCOMINE database analysis. Increased levels of sTNFRII reduced the risk of subcentimetre lung adenocarcinoma by 89% (P < .001). Patients with a higher level of BLC had a 2.70-fold (P < .01) higher risk of subcentimetre adenocarcinoma. Furthermore, a higher BLC/TNFRII ratio was related to a 35-fold higher risk of subcentimetre adenocarcinoma. TNFRII showed good specificity, sensitivity and accuracy (0.72, 0.75 and 0.73, respectively), with an AUC of 0.73 (P < .001). In conclusion, the present study assessed the value of sTNFRII as a potential biomarker to predict the risk of subcentimetre lung adenocarcinoma and provided evidence for the further use of TNFRII as an auxiliary marker in the diagnosis of subcentimetre lung adenocarcinoma.     

低氧促进肺腺癌A549细胞迁移的机制*

目的: 探讨低氧促进肺腺癌A549细胞迁移的机制。方法: 培养肺腺癌A549细胞,转染慢病毒获得稳定敲低ACC1的A549细胞株,转染si-RNA获得敲低SREBP-1的A549细胞。分别以低氧(5％ O₂)联合低氧诱导因子1α(HIF-1α)抑制剂PX-478(25 μmol)处理A549细胞,低氧联合亚油酸(LA)(20 μmol)处理敲低ACC1的A549细胞,低氧处理敲低胆固醇调节原件结合蛋白1(SREBP-1)的A549细胞。Transwell实验检测细胞迁移,蛋白质印迹法检测HIF-1α、ACC1及上皮-间质转化(EMT)相关波形蛋白(Vimentin)及E-钙黏蛋白(E-Cadherin)的表达与SREBP-1的表达,实时荧光定量聚合酶链反应(RT-qPCR)检测低氧联合HIF-1α抑制剂PX-478(25 μmol)处理A549细胞后ACC1及SREBP-1 mRNA水平变化。每项实验重复三次。结果: 与常氧组相比,低氧组A549细胞迁移数增加,ACC1与HIF-1α表达上调(P均＜0.01),SREBP-1表达上调(P＜0.05);与低氧对照组相比,PX-478(25 μmol)抑制A549细胞迁移,SREBP-1表达下调(P＜0.05);低氧处理A549细胞后ACC1 mRNA上升(P＜0.05),SREBP-1 mRNA水平上升(P＜0.01);低氧并使用PX-478(25 μmol)处理A549细胞24 h,ACC1 mRNA水平下降(P＜0.05),SREBP-1 mRNA 水平下降(P＜0.01);转染si-RNA获得敲低SREBP-1的A549细胞,Transwell 实验显示si-SREBP-1组细胞迁移数较常氧对照组减少(P＜0.01);低氧处理si-SREBP-1组与si-NC组,与对照组相比si-SREBP-1组细胞迁移数减少(P＜0.01)但与常氧组相比差异无统计学意义(P＞0.05);Western blot检测到si-SREBP-1组ACC1表达较对照组下降(P＜0.01);低氧处理si-SREBP-1组,ACC1表达较对照组下降(P＜0.01);敲低ACC1抑制A549细胞迁移(P＜0.05),敲低ACC1后A549细胞在常氧和5％ O₂条件下细胞迁移数目差异无统计学意义(P＞ 0.05);低氧处理敲低ACC1的A549细胞并给予LA(25 μmol)促进A549细胞迁移(P＜0.05)。结论: 低氧通过HIF-1α/SREBP-1/ACC1途径调节脂肪酸代谢进而促进肺腺癌A549细胞迁移。     

Association of glutathione S-transferase P1 gene polymorphism with the risk of small-cell carcinoma of lung cancer

Abstract

The conclusions of the published reports on the relationship between glutathione S-transferase P1 (GSTP1) gene polymorphism and the risk of small-cell carcinoma of lung cancer are still debated. GSTP1 is one of the important mutant sites reported at present. This meta-analysis was performed to evaluate the association between GSTP1 and the risk of small-cell carcinoma of lung cancer. The association investigations were identified from PubMed and Cochrane Library, and eligible studies were included and synthesized using meta-analysis method. Ten reports were included into this meta-analysis for the association of GSTP1 A/G gene polymorphism and small-cell carcinoma of lung cancer. The G allele and GG genotype were not associated with the susceptibility of risk of small-cell carcinoma in overall populations, East-Asians and Turkish population. However, there was an association between GG genotype with the risk of small-cell carcinoma in Caucasians. In conclusion, GG genotype was associated with the risk of small-cell carcinoma in Caucasian patients with lung cancer. However, GSTP1 A/G gene polymorphism is not associated with the susceptibility of small-cell carcinoma in overall populations, East-Asians and Turkish population.     

LncRNA TTN-AS1 drives invasion and migration of lung adenocarcinoma cells via modulation of miR-4677-3p/ZEB1 axis

Lung adenocarcinoma is the most prevalent type of lung cancer with a high incidence and mortality worldwide. Metastasis is the major cause of high death rate in lung cancer and the potential mechanism of lung adenocarcinoma metastasis remains indistinct. Emerging investigations have demonstrated that long noncoding RNA is a kind of non–protein coding RNA and plays a critical role in cancer progression and metastasis. TTN antisense RNA 1 (TTN-AS1) has been reported to promote cell growth and metastasis in cancer. However, the function of TTN-AS1 in lung adenocarcinoma is still to be illustrated. In this study, we observed that TTN-AS1 was upregulated in tissues and cells of lung adenocarcinoma and associated with poor overall survival. TTN-AS1 promoted cell proliferation, migration, invasion, and epithelial-mesenchymal transition in lung cancer. TTN-AS1 directly bound with miR-4677-3p and negatively regulated miR-4677-3p. MiR-4677-3p rescued the inhibitive impacts of TTN-AS1 knockdown on lung adenocarcinoma. Furthermore, zinc finger E-box binding homeobox 1 (ZEB1) was the target of miR-4677-3p, and TTN-AS1 modulated ZEB1 by competing for miR-4677-3p. TTN-AS1 drove the invasion and migration of lung adenocarcinoma cells by targeting the miR-4677-3p/ZEB1 axis. To sum up, our study offers insights into the mechanism of TTN-AS1 in lung adenocarcinoma metastasis and targeting the TTN-AS1/miR-4677-3p/ZEB1 axis may be the potential innovate therapeutic strategy for the patients with lung adenocarcinoma.