Human immunodeficiency virus Tat‐TIP30 interaction promotes metastasis by enhancing the nuclear translocation of Snail in lung cancer cell lines

Lung cancer patients with human immunodeficiency virus (HIV) have a poorer prognosis than do patients without HIV infection. HIV1 Tat is a secreted viral protein that penetrates the plasma membrane and interacts with a number of proteins in non‐HIV‐infected cells. The loss of function of Tat‐interacting protein 30 (TIP30) has been linked to metastasis in non‐small cell lung cancer (NSCLC). However, it is unknown how the interaction of HIV1 Tat with TIP30 regulates the metastasis of NSCLC cells. In this study, the overexpression of TIP30 decreased tumor growth factor‐β‐induced epithelial‐to‐mesenchymal transition (EMT) and invasion of NSCLC cells, whereas the knockdown of TIP30 promoted EMT, invasion and stemness. Exposure to recombinant HIV1 Tat proteins promoted EMT and invasion. A mechanistic study showed that the interaction of HIV1 Tat with TIP30 blocked the binding of TIP30 to importin‐β, which is required for the nuclear translocation of Snail. Indeed, the loss of TIP30 promoted the nuclear translocation of Snail. In vivo studies demonstrated that the overexpression of TIP30 inhibited the metastasis of NSCLC cells. In contrast, the coexpression of HIV1 Tat and TIP30 diminished the inhibitory effect of TIP30 on metastasis. Immunohistochemistry confirmed that TIP30 overexpression reduced the nuclear localization of Snail, whereas the coexpression of HIV1 Tat and TIP30 increased nuclear Snail in metastatic tumors. In conclusion, the binding of HIV1 Tat to TIP30 enhanced EMT and metastasis by regulating the nuclear translocation of Snail. Targeting Tat‐interacting proteins may be a potential therapeutic strategy to prevent metastasis in NSCLC patients with HIV infection.     

MicroRNA-328 is associated with (non-small) cell lung cancer (NSCLC) brain metastasis and mediates NSCLC migration

Brain metastasis (BM) can affect ～ 25% of nonsmall cell lung cancer (NSCLC) patients during their lifetime. Efforts to characterize patients that will develop BM have been disappointing. microRNAs (miRNAs) regulate the expression of target mRNAs. miRNAs play a role in regulating a variety of targets and, consequently, multiple pathways, which make them a powerful tool for early detection of disease, risk assessment, and prognosis. We investigated miRNAs that may serve as biomarkers to differentiate between NSCLC patients with and without BM. miRNA microarray profiling was performed on samples from clinically matched NSCLC from seven patients with BM (BM+) and six without BM (BM-). Using t-test and further qRT-PCR validation, eight miRNAs were confirmed to be significantly differentially expressed. Of these, expression of miR-328 and miR-330-3p were able to correctly classify BM+ vs. BM- patients. This classifier was used on a validation cohort (n = 15), and it correctly classified 12/15 patients. Gene expression analysis comparing A549 parental and A549 cells stably transfected to over-express miR-328 (A549-328) identified several significantly differentially expressed genes. PRKCA was one of the genes over-expressed in A549-328 cells. Additionally, A549-328 cells had significantly increased cell migration compared to A549 cells, which was significantly reduced upon PRKCA knockdown. In summary, miR-328 has a role in conferring migratory potential to NSCLC cells working in part through PRKCA and with further corroboration in additional independent cohorts, these miRNAs may be incorporated into clinical treatment decision making to stratify NSCLC patients at higher risk for developing BM.     

MicroRNA-195 inhibits non-small cell lung cancer cell proliferation,migration and invasion by targeting MYB

MicroRNA-195 (miR-195) has been implicated in several other cancers; however, its role in non-small cell lung cancer (NSCLC) remains unclear. In this study, we demonstrated that miR-195 was significantly down-regulated in NSCLC samples and cell lines compared with corresponding normal counterparts. In vitro and in vivo functional assays demonstrated that modulation of miR-195 expression affected NSCLC cell proliferation, migration and invasion. Using miRNA target prediction algorithms and reporter assays, we demonstrated that miR-195 suppressed the expression of MYB both at the mRNA and protein level, and was directly bound to the 3′untranslated region of MYB mRNA. Overexpression of MYB in NSCLC cells using an ectopic expression vector restored the decreased cell proliferation, migration and invasion effects induced by miR-195. Finally, we observed an inverse correlation between MYB and miR-195 in NSCLC. Taken together, our findings indicated that miR-195 functions as tumour suppressor in NSCLC, and the miR-195/MYB axis might represent a potential therapeutic target for NSCLC intervention.     

E-cadherin在非小细胞肺癌中的表达及临床意义

目的探讨E-cadherin在非小细胞肺癌(NSCLC)中的表达.方法采用免疫组织化学SABC法,对常规10%福尔马林固定、石蜡包埋的NSCLC90例分别进行Ecadherin的检测.结果 E-cadherin在NSCLC中表达阳性率为52.2%(47/90).在NSCLC中E-cadherin与PTNM分期、病理分级、淋巴结转移及预后有显著相关性,与病理类型无关.结论 E-cadherin缺失可促使肿瘤转移,可能是NSCLC转移的重要因素之一,可作为判断NSCLC转移及预后有价值的指标之一.     

新型雌激素受体 GPR30在非小细胞肺癌中的表达

目的：研究 GPR30与 ERα、ERβ、HER2、HER3在非小细胞肺癌中的表达及相关性,并分析 GPR30和ERβ与临床病理特征之间的关系。方法：采用免疫组织化学方法检测60例术后非小细胞肺癌组织样本中GPR30、ERα、ERβ、HER2、HER3的表达。结果：GPR30表达在有淋巴结转移、腺癌、低分化、III 期肿瘤中明显高于无淋巴结转移、鳞癌、中高分化、I - II 期肿瘤,差异有统计学意义（P <0.05）。ERβ表达在腺癌中显著高于鳞癌（P <0.05）。GPR30表达与 ERβ和 HER3呈中度正相关（r =0.607,P =0.000;r =0.510,P =0.000）。结论：GPR30与 ERβ或 HER2- HER3的信号途径可能存在相关性,共同参与非小细胞肺癌的发生发展。     

Role of skip metastasis to mediastinal lymph nodes in non-small cell lung cancer

BACKGROUND: Skip metastasis to mediastinal lymph nodes is a well-known phenomenon in non-small cell lung cancer (NSCLC). Little is reported in the literature about its clinical importance. It is still under discussion whether any prognostic differences exist between resected NSCLC with either skip metastases or continuous mediastinal lymph node metastases (N2). PATIENTS AND METHODS: We analyzed retrospectively the data of 45 patients with a pN2-stage, who underwent resection for NSCLC. Seventeen of these patients (37.8%), showing no metastatic involvement of hilar (N1) lymph nodes, were compared to the remaining 28 patients with infiltration of hilar nodes (N1) as well as N2 nodes. RESULTS: Multivariate analysis showed no statistically significant difference between the skip metastasis and the continuous N2 group regarding sex, age, histology, T- or M-status. The frequency of skip metastasis was higher in patients with a primary tumor in the upper lobe (n = 12, 71%) compared to the lower lobe (n = 5, 29%). This difference was not statistically significant. In patients with a non-continuous lymph node spread, 29 out of 119 resected mediastinal lymph nodes were infiltrated (1.7 per patient, range: 1-10). Compared to 83 metastatic involved lymph nodes out of 198 resected mediastinal nodes (three per patient, range: 1-10) in patients with involvement of N1 and N2 nodes (P = 0.034, Mann-Whitney test). The 5-year survival rate of pN2 patients with skip metastasis was 41% compared to 14% in patients with involvement of N1 and N2 nodes (P = 0.019). CONCLUSIONS: pN2 patients with mediastinal lymph node skip metastasis have a more favorable prognosis compared to pN2 patients with continuous infiltration of the regional lymph nodes. Patients with a continuous lymph node involvement show an increased number of infiltrated mediastinal lymph nodes per patient compared to patients with a non-continuous spread. Skip metastasis is an independent prognostic factor of survival. The presence of skip metastasis seems to be a unique subgroup of pN2 disease in NSCLC.     

Akt1 inhibition by RNA interference sensitizes human non-small cell lung cancer cells to cisplatin

Akt/protein kinase B signaling is very important for cancer cell survival and growth when cells are exposed to various apoptotic stimuli. Akt is constitutively activated in NSCLC cells and is a potential target for enhancing the cytotoxicity of chemotherapeutic agents in treatment of NSCLC. In our study, we investigated whether down-regulating Akt1 using RNAi techniques can enhance sensitivity to cisplatin in NSCLC cells. An siRNA targeting Akt1 significantly decreased the protein level of Akt1 and the activity of ERK. Treatment of these cells with 20 muM cisplatin increased apoptotic cell death approximately 2.6-fold compared to cells transfected with a scrambled siRNA. While Akt activity was slightly reduced, ERK activity was greatly increased in cells treated with cisplatin alone. Pretreatment of these cells with the selective MEK inhibitor U0126 effectively reduced the level of cisplatin-induced apoptosis. These results imply that cisplatin-induced MEK/ERK activation appears to mediate apoptotic cell death, but that constitutively activated Akt1 and/or ERK pathway may mediate resistance to cisplatin in NSCLC cells. Taken together, our data demonstrate that down-regulation of Akt1 using RNAi enhances the chemosensitivity of NSCLC cells to cisplatin.     

Expression of telomeric repeat binding factor 1 in non-small cell lung cancer

BACKGROUND AND OBJECTIVES: Telomeric repeat binding factor 1 (TRF1) is crucial for forming and maintaining the protective telomeric structure. However, the relationship between TRF1 and non-small cell lung cancer(NSCLC) is not well understood. With this background, we investigated the expressions of the mRNA encoded by the TRF1 gene in cancer tissue and the paired non-cancerous tissue. We also examined whether TRF1 expression is correlated with histopathological features. METHODS: From October 2004 to August 2005, 40 patients with NSCLCs had undergone curative operations, including 29 males and 11 females. There were 20 cases of squamous cell carcinoma and 20 cases of adenocarcinoma. We measured the expression of TRF1 mRNA using RT-PCR on 40 surgically resected specimens and the paired non-cancerous tissues. RESULTS: TRF1 mRNA was significantly downregulated in cancer tissue compared with the paired non-cancerous tissue. Additionally, the expression of TRF1 mRNA was significantly associated with the grade of tumor differentiation. No significant difference of TRF1 mRNA level was found between sexes, or among different T-status, clinical stages, pathological subtypes, and lymph node metastasis. CONCLUSIONS: Downregulation of TRF1 mRNA expression appeared in lung cancer tissue. TRF1 may play a significant role in cell differentiation in NSCLC.     

DKK1 促进非小细胞肺癌线性程序性坏死和血管生成拟态形成*

目的:探讨非小细胞肺癌（non-small cell lung cancer ,NSCLC ）中DKK 1 影响线性程序性坏死（linearly patterned programmed cellnecrosis ,LPPCN ）和血管生成拟态（vasculogenic mimicry ,VM）的机制。方法:收集人NSCLC 标本173 例,H&E染色检测LP?PCN ,CD31/PAS 双染检测VM,免疫组织化学检测DKK 1 及相关蛋白表达,分析其临床病理意义及相互关系,进而裸鼠H 460-DKK 1 移植瘤体内验证研究假设。结果:NSCLC 中14.45%（25/ 173）存在VM,49.71%（86/ 173）具有LPPCN ,LPPCN （+）组25.6%（22/86）形成VM,二者均与分化差、TNM 分期晚、易复发转移和预后差相关。VM（+）组和LPPCN （+）组DKK 1 均高表达,均与VE-cad?herin、MMP-2、β -catenin 核表达及Twist1 正相关。H 460-DKK 1 移植瘤模型证实DKK 1 促进VM和LPPCN 及相关蛋白表达上调。结论:DKK 1 引起的β -catenin、Twist1 表达上调可促进NSCLC 中LPPCN 和VM形成。      

局部晚期非小细胞肺癌的放射治疗

本文综述了近三年来在局部晚期非小细胞肺癌放射治疗方面的临床研究进展。①术前的诱导治疗，包括术前的化疗或术前放化疗综合应用。②能手术切除病人的化放疗综合治疗，提出了除手术治疗外，综合使用化疗和放射治疗是这些病人的另一个选择。③不能手术切除病人的化疗和放疗。强调了全身化疗的重要性，讨论了化放综合治疗的模式及其疗效和治疗并发症。④三维适形放疗，已有的文献支持该方法能较好地保护肺组织，从而提高肿瘤的照射剂量。因此肿瘤的局控率和生存率均有明显改善。⑤非常规分割放疗。加速超分割放疗在一个较短的照射时间内给一个相对高的放射剂量，结果显示提高了局控率。因而有继续研究的前景。⑥放疗保护剂的试用。Amifostine已被用于肺癌的放疗，实践证实能减少放射性肺炎和食管炎的产生。     

MicroRNA‐331‐3p inhibits epithelial‐mesenchymal transition by targeting ErbB2 and VAV2 through the Rac1/PAK1/β‐catenin axis in non‐small‐cell lung cancer

MicroRNAs have been reported to play critical roles in the regulation of non‐small‐cell cancer (NSCLC) development, but the role of microRNA (miR)‐331‐3p in NSCLC is still unclear. In this study, the expression levels of miR‐331‐3p in NSCLC tumor tissues and adjacent normal tissues were examined by quantitative RT‐PCR, and the relationship between miR‐331‐3p expression and patient clinicopathological characteristics was analyzed. The effects of miR‐331‐3p on epithelial‐mesenchymal transition (EMT), migration, and metastasis of NSCLC cells were determined in vitro and vivo. Direct functional targets of miR‐331‐3p were identified by luciferase reporter assay, western blot assay, immunohistochemical staining, and rescue assay. The downstream pathway regulated by miR‐331‐3p was identified by immunofluorescence, immunoprecipitation, and Rac1 activity examination. Our results showed that miR‐331‐3p was significantly downregulated in NSCLC tumor tissues and was correlated with clinicopathological characteristics, and miR‐331‐3p could be an independent prognostic marker for NSCLC patients. Furthermore, miR‐331‐3p significantly suppressed EMT, migration and metastasis of NSCLC cells in vitro and in vivo. Both ErbB2 and VAV2 were direct functional targets of miR‐331‐3p. The activities of Rac1, PAK1, and β‐catenin were regulated by miR‐331‐3p through ErbB2 and VAV2 targeting. These results indicated that miR‐331‐3p suppresses EMT, migratory capacity, and metastatic ability by targeting ErbB2 and VAV2 through the Rac1/PAK1/β‐catenin axis in NSCLC.     

Randomized phase 2 sequencing and pharmacokinetic study of gemcitabine and oxaliplatin in advanced non-small cell lung cancer

Aim: This multicentre phase II trial examined the combination of gemcitabine and oxaliplatin in patients with advanced non‐small cell lung cancer (NSCLC). The effect of sequence administration was randomized and pharmacokinetics (PK) assessed. Methods: Eligible patients had stage IIIB or IV or recurrent NSCLC, no prior chemotherapy, World Health Organization performance status ≤2 and measurable disease. Treatment comprised: gemcitabine (1250 mg/m²) and oxaliplatin (70 mg/m²), each given on days 1 and 8 of a 21‐day cycle. Patients were randomized 1:1 to the sequencing of the two drugs for the duration of their treatment. The primary end‐point was response rate (RR). Secondary end‐points included progression‐free survival (PFS), overall survival (OS), toxicity, PK and the effect of drug sequencing. Results: A total of 46 patients were enrolled of whom 43 were evaluable for response. Overall 13 patients (30%) achieved a partial response, PFS was 4.2 months (95% CI 2.8–5.8 months), and OS was 6.8 months (95% CI 4.4–10.1 months). There was only one case of grade 3 neurosensory toxicity despite a median cumulative oxaliplatin dose in excess of 500 mg/m². No differences in clinical or PK end‐points were observed between the two different sequencing arms. Conclusion: This oxaliplatin and gemcitabine schedule has shown activity in advanced NSCLC with modest toxicity. Neither clinical nor PK outcomes were influenced by the sequencing of these agents, although definite conclusions are limited by small patient numbers. The favorable toxicity profile of this doublet, in light of an encouraging RR, warrants its further investigation in NSCLC.