Proteomics-based identification of tumor relevant proteins in lung adenocarcinoma

Background

Lung cancer has the highest mortality rate among malignant tumors. Proteomics is a powerful tool to identify protein biomarkers. The identification of protein biomarkers associated with lung adenocarcinoma would have significance for making prognoses and designing targeted therapies.

Methods

In our study, we applied a two-dimensional difference gel electrophoresis approach coupled to a matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis for the identification of proteins differentially expressed between lung adenocarcinoma and the paired normal bronchial epithelial tissues derived from seven patients (four of them developed distant metastasis after operation). In addition, we chose two candidate proteins and examine their expression levels in lung adenocarcinoma and adjacent normal tissues using immunohistochemistry methods, and their expression levels in serum of patients and healthy donors by ELISA.

Result

In this study, 173 proteins were found to be differentially expressed (ratio > 1.5 or < –1.5, P ≤ 0.05), and 22 of them were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Thirteen proteins were at lower levels in the lung adenocarcinoma group, while nine proteins were at higher abundance. Immunohistochemistry analysis confirmed the expression levels of the two candidate proteins. The differential expression of the candidate secreted protein in serum from lung adenocarcinoma samples and healthy controls was showed by ELISA.

Conclusion

Our results demonstrated a differential protein expression pattern for lung adenocarcinoma compared with the paired normal bronchial epithelial tissues. Further functional validation of candidate proteins is ongoing and might provide new insights in lung adenocarcinoma.     

Guanylate‐binding protein 1 correlates with advanced tumor features,and serves as a prognostic biomarker for worse survival in lung adenocarcinoma patients

ObjectiveGuanylate‐binding protein 1 (GBP1) is reported to promote tumor progression and treatment resistance in lung cancer, and presents as a prognostic biomarker in several solid tumors. However, the related research of GBP1 in clinical management of lung adenocarcinoma is still lacking. Therefore, the present study aimed to detect the clinical role of GBP1 in lung adenocarcinoma.MethodsThe clinical data of 221 lung adenocarcinoma patients were retrospectively analyzed, and then, their tumor tissue specimens and paired adjacent tissue specimens were retrieved for GBP1 detection via immunohistochemistry (IHC) assay.ResultsGBP1 expression was upregulated in tumor tissues compared with adjacent tissues (P < .001). Moreover, high tumor GBP1 expression was associated with larger tumor size (P = .030), positive lymph node (LYN) metastasis (P = .001), advanced TNM stage (P = .001), and abnormal preoperative carcinoembryonic antigen (CEA) level (P = .026). Furthermore, tumor GBP1 high expression was correlated with reduced disease‐free survival (DFS) and overall survival (OS), and was of independent value in predicting worse DFS and OS. Additionally, data analysis of 1144 lung cancer patients derived from KMplot database (www.kmplot.com) further verified that GBP1 expression was negatively correlated with OS (P = .009).ConclusionGBP1 correlates with advanced tumor features and worse survival profiles, suggesting its value to be a prognostic biomarker in management of lung adenocarcinoma.     

Expression and clinical significance of tyrosine phosphatase SHP-2 in colon cancer

Protein-tyrosine phosphatase SHP-2, encoded by gene PTPN11, has been identified as a tumor-promoting factor in several types of leukemia and is hyper-activated by other mechanisms in some solid tumors including gastric cancer, breast cancer, non-small cell lung cancer (NSCLC), etc. But few were reported on the expression and significances of SHP-2 in colon cancer. Here, we detect SHP-2 expression in colon cancer cells, colon cancer-induced by AOM + DSS in mice and 232 human colon cancer specimens, including 58 groups of self-matched adjacent peritumor tissues and normal tissues. We found that compared to the normal colon tissues, SHP-2 significantly decreased in tumor tissues (P < 0.001). The same results were got in colon tumor cells as well as mice colon tumors. And in humans samples, low SHP-2 expression showed a significantly correlation with poor tumor differentiation (P < 0.05), late TNM stage (P = 0.1666) and lymph node metastasis (P < 0.05)     

HES5蛋白与肺腺癌患者病情及预后的相关性研究

目的探究HES5蛋白表达水平与肺腺癌患者病情及预后相关性。方法回顾性收集江苏省中医院心胸外科离体后30 min内立即冻存于液氮中的4组肺腺癌组织和癌旁组织,采用Western Blot法分析HES5表达特征;收集2017年4月至2019年4月本院切除并经病理证实的48例肺腺癌标本(其中距肿瘤边缘>5 cm组织为癌旁正常组织),采用免疫组化染色法检测癌组织和癌旁组织HES5蛋白蛋白表达量,分析HES5蛋白表达量与性别、年龄、身体质量指数(BMI)、肿瘤直径、分化程度、肿瘤转移、肿瘤分期相关性;获取由手术时间起至患者死于癌症及癌症相关事件或纳入本研究36个月止的临床资料,根据患者生存情况分为生存组和死亡组,分析影响肺腺癌预后影响因素。结果HES5蛋白在分子量约为18 k Da处有显著表达,HES5蛋白在肺腺癌组织中表达升高,HES5蛋白在癌旁正常组织中表达降低,两者比较差异有统计学意义(P<0.05);肺腺癌组织中HES5阳性表达率高于癌旁组织中HES5阳性表达率(P<0.05)。TNM分期Ⅱ/Ⅲ期、有淋巴结转移、肿瘤直径>3 cm的肺腺癌患者HES5蛋白表达量高于分期I期、无淋巴结转移、肿瘤直径≤3 cm的肺腺癌患者HES5蛋白表达量(P<0.05)。3年随访期间存活患者28例(占比48.28%),死亡30例(占比51.72%);肿瘤直径>3 cm、分化程度为低/中分化、有淋巴结转移、TNM分期Ⅱ/Ⅲ期、HES5蛋白高表达的肺腺癌患者存活率低于肿瘤直径≤3 cm、分化程度为高分化、无淋巴结转移、TNM分期I期、HES5蛋白低表达的肺腺癌患者存活率(P<0.05)。COX多因素结果显示,HES5蛋白表达、TNM分期是影响肺腺癌患者预后独立影响因素(P<0.05)。结论HES5蛋白在肺腺癌肿瘤组织中呈高表达水平,可负向调控肺腺癌患者肿瘤分级、肿瘤分期、肿瘤转移及病情进展,检测HES5蛋白表达水平可为肺腺癌患者早期诊疗及预后评估提供参考依据。     

Bcl-2 gene silence enhances the sensitivity toward 5-Fluorouracil in gastric adenocarcinoma cells

Because of increased insensitivity or resistance to chemical treatment in tumor patients, specific apoptotic gene silence may provide a rational approach for the development of novel therapeutic strategies. This study was to investigate whether downregulation of Bcl-2 expression by small interfering RNA (siRNA) against the Bcl-2 gene would enhance the apoptosis and sensitivity of gastric adenocarcinoma SGC-7901 cell to 5-Fluorouracil. Transfections of SGC-7901 cells with siRNA were performed using cationic liposomes. Sequence-specific downregulation of Bcl-2 expression was measured by RT-PCR and Western blot analysis. Cell proliferation assay was determined by MTT assay and apoptotic cell rates were determined by flow cytometry assay. Results showed that the siRNA could downregulate Bcl-2 expression, which increased apoptosis and sensitivity of SGC-7901 cell to 5-Fluorouracil (P < 0.05). This study indicated that inhibition of Bcl-2 expression by siRNA would be useful a new useful protocol to increase the effect of 5-Fluorouracil on treatment of gastric adenocarcinoma, which may play an important role in developing novel therapeutic strategies in the future.     

Inhalation delivery of Telmisartan enhances intratumoral distribution of nanoparticles in lung cancer models

The purpose of the present study was to evaluate the effect of Telmisartan (Tel) and Losartan (Los) on nanoparticle intratumoral distribution and anticancer effects in lung cancer. A549 lung tumor cells were orthotopically and metastatically administered to Nu/nu mice. Fluorescent polystyrene nanoparticles (FPNPs, size ~ 200 nm) beads were used to study their intratumoral distribution after Tel and Los treatments. Animals were administered with FPNPs and after 2 h, FPNPs intratumoral distribution was studied by fluorescent microscopy. Tel (~ 1.12 mg/kg) and Los (~ 4.5 mg/kg) were administered by inhalation delivery at alternative days for 4 weeks to tumor bearing animals. Collagen-1, transforming growth factor beta 1 (TGF-β1), cleaved caspase-3, Vimentin and E-Cadherin expressions were studied by western blotting. To correlate the AT₁ receptor blockage to anticancer effects, VEGF levels and microvessel densities (MVD) were quantified. Los and Tel treated group resulted in the 5.33 and 14.33 fold increase respectively in the FPNPs intratumoral distribution as compared to the controls. Tel treatment attenuated 2.23 and 1.70 fold Collagen 1 expression compared to untreated control and Los groups, respectively. Further, in Tel and Los treated groups, the TGF-β1 active levels were significantly (p < 0.05) decreased. Tel (at four times less dose) was 1.89 and 1.92 fold superior in anticancer activity to Los respectively in A549 orthotopic and metastatic tumor models (p < 0.05) when given by inhalation route. Tel, by virtue of its dual pharmacophoric nature could be an ideal candidate for combination therapy to improve the nanoparticle intratumoral distribution and anticancer effects.     

TPX2 siRNA regulates growth and invasion of esophageal cancer cells

Purpose

Observe how specific small RNA interference (siRNA) aimed at TPX2 gene suppresses TPX2 gene expression in esophageal cancer EC9706 cells and the effect on esophageal cancer cell growth and invasion ability.

Methods

Transfect TPX2 siRNA into EC9706 cells via lipofectamin 2000. The experiments were divided into three groups, a negative control, a blank control and an siRNA interference group (24 h, 48 h, 72 h, 96 h). We examined RNA and protein level alteration of the TPX2 gene after TPX2 siRNA transfection by RT-PCR and Western blot analysis. Detection of how TPX2 siRNA influences EC9706 cell proliferation was done by MTT, cell apoptosis monitored through Tunel assay, in vitro invasion ability via Boyden chamber and cell cycle change by flow cytometry.

Results

After effective siRNA transfection, TPX2 mRNA and protein expression level in siRNA interference group were (0.31 ± 0.08, 0.39 ± 0.12),72 h after transfection, significantly lower than blank control group (1.00 ± 0.01) and negative control group (0.98 ± 0.11), (F = 71.182, t1 = 8.17, t2 = 7.90, P < 0.05); MTT results demonstrated that cell growth and proliferation were inhibited and the inhibition rate was up to 35.4% (P < 0.05) compared with the control group. TUNEL results indicated that cell apoptosis index in siRNA interference group was 18.28 ± 0.35, higher than that in blank control group (4.07 ± 0.26)and negative control group (4.13 ± 0.22), (F = 244.5, t1 = 60.61, t2 = 53.32, P < 0.01). Boyden chamber results showed that the transmembrane cell number was 45.30 ± 8.08 in siRNA interference group, less than blank control group (121.90 ± 7.83), (F = 122.46, t1 = 11.81, t2 = 10.47, P < 0.01); besides, in siRNA interference group cell invasion inhibition rate was 71.42 ± 9.12, higher than negative control group (5.65 ± 3.55), (t = 14.256, P < 0.01). Flow cytometry results illustrated that more EC9706 cells went into apoptosis and cell cycle arrested in S phase. Similar results were obtained by in vivo transplantation, as TPX2 siRNA transfection significantly reduced tumor growth of the xenograft in nude mice.

Conclusion

siRNA could effectively inhibit the invasion and metastasis of EC9706 cells, promote the apoptosis of tumor cells and may become a new approach for treatment of esophageal carcinoma.     

Stage dependent destructuration of neuro-endocrine-immune system components in lung cancer patients

Background

Close relationships among the nervous, endocrine and immune system components maintain body homeostasis. Alteration of time-related prophile of variation of system components and loss of integrated function may favour the developing of cancer and may be aggravated in the presence of neoplastic disease. The aim of our study was to evaluate the prophiles of time-related variation of neuro-endocrine-immune system components in lung cancer patients.

Methods

Peripheral blood samples were collected at intervals of 4 hours for 24 hours from 11 healthy subjects (age range 35-53 years, mean age ± s.e. 43.6 ± 1.7) and nine patients suffering from nonsmall cell lung cancer (age range 43-63 years, mean age ± s.e. 51.0 ± 2.4). In each blood sample, lymphocyte subpopulations (CD3, CD4, CD8, HLA-DR, CD16, CD20, CD25, γδTcR) were analyzed and melatonin, cortisol, TRH, TSH, free thyroxine, GH, IGF1 and interleukin IL2 on serum were measured.

Results

In our I-II stage lung cancer patients CD8+ lymphocytes (P = 0.01), and in particular the T suppressor subset (P < 0.0001), CD20+ cells (P = 0.05), γδTCR expressing cells (P < 0.01) and IGF1 (P = 0.004) were diminished, whereas CD16+ cells (P < 0.0001), CD25+ cells (P = 0.03), free thyroxine (P = 0.001) and GH (P < 0.0001) were increased in respect of healthy subjects. In our III-IV stage lung cancer patients CD8+ lymphocytes (P = 0.003) and in particular the T suppressor subset (P < 0.0001), CD20+ cells (P = 0.05), γδTCR expressing cells (P = 0.01), melatonin (P = 0.03), TSH (P = 0.006) and IGF1 (P < 0.0001) were diminished, whereas CD4+ cells (P = 0.002), CD16+ cells (P < 0.0001), CD25+ cells (P = 0.002), cortisol (P = 0.003), TRH (P = 0.004), free thyroxine (P = 0.001), GH (P < 0.0001) and IL2 (P = 0.0002) were increased in respect of healthy subjects. A statistically significant difference was evidenced between the two groups of cancer patients for the values of CD16+ cells (P < 0.0001), free thyroxine (P = 0.001) and IGF1 (P < 0.0001) higher in I-II stage lung cancer patients and for the values of CD4+ cells (P < 0.0001), γδTCR expressing cells (P = 0.002), TRH (P = 0.002) and IL2 (P = 0.01) higher in III-IV stage lung cancer patients. Lung cancer patients showed alteration of the pattern of circadian variation of CD3+, CD8+, CD8+ dim, CD16+, CD20+ and γδTCR expressing cells and of cortisol, TSH and GH serum levels. Pair-wise comparisons showed severe and stage dependent alterations in lung cancer patients.

Conclusions

The prophiles of time-related variation of neuro-endocrine-immune system components are altered in a stage dependent manner in lung cancer patients and this alteration may impair the customary integrated system function.     

Is leptin a predictive factor in patients with lung cancer?

Objectives

The aim of this study was to evaluate the expression and clinical significance of leptin in lung cancer.

Methods

126 patients with lung cancer ranged from 30 to 83 years of age were studied. Serum leptin levels were determined by ELISA. The mRNA and protein levels of leptin in normal and lung cancer tissues were measured by RT-PCR and immunohistochemistry. The relationships between leptin levels and clinicopathological factors were evaluated by Wilcoxon rank sum or Kruskal–Wallis H test.

Results

Serum leptin levels in lung cancer patients were significantly higher compared to those in controls and leptin expression in lung cancer tissue was markedly increased than that in normal lung tissue (both P < 0.050).

Conclusions

Determination of leptin levels might provide useful predictive information for lung cancer.     

PRAME mRNA和SOX9 mRNA在肺腺癌中的表达及临床意义

目的探讨黑色素瘤特异性抗原(PRAME)mRNA和性别决定基因盒9(SOX9)mRNA在肺腺癌中的表达及临床意义。方法选取2015年2月至2018年4月于该院接受手术治疗的73例肺腺癌患者为研究对象,术中收集肺腺癌癌组织及癌旁正常组织,采用实时荧光定量PCR检测不同组织中PRAME mRNA、SOX9 mRNA的相对表达水平并进行比较;分析肺腺癌患者癌组织PRAME mRNA、SOX9 mRNA表达与临床病理特征的关系;采用Spearman相关分析PRAME mRNA表达与SOX9 mRNA表达的相关性;分析PRAME mRNA、SOX9 mRNA表达与肺腺癌患者预后的关系;采用COX回归模型分析影响肺腺癌患者预后的因素。结果与癌旁正常组织相比,肺腺癌癌组织PRAME mRNA的相对表达水平降低(P<0.05),SOX9 mRNA的相对表达水平升高(P<0.05)。PRAME mRNA、SOX9 mRNA表达与肺腺癌患者性别、年龄、肿瘤大小无关(P>0.05),与病理分级、临床分期、淋巴结转移有关(P<0.05)。Spearman相关分析显示,肺腺癌癌组织PRAME mRNA表达与SOX9 mRNA表达呈负相关(P<0.05)。与预后良好组相比,预后不良组肺腺癌患者PRAME mRNA的相对表达水平降低(P<0.05),SOX9 mRNA的相对表达水平升高(P<0.05)。COX回归模型分析结果显示,PRAME mRNA低表达、SOX9 mRNA高表达、低分化、临床分期Ⅲ期、有淋巴结转移均是影响肺腺癌患者预后的独立危险因素(P<0.05)。结论肺腺癌癌组织中PRAME mRNA、SOX9 mRNA表达异常,且两者呈负相关,可能为肺腺癌的病情及预后评估提供参考依据。     

PITX1基因在肺腺癌中的表达及临床预后意义

目的:探讨同源域转录因子1(paired like homeodomain 1,PITX1)基因在肺腺癌组织和正常组织中的表达水平及其与临床病理特征和预后之间的相关性。方法:综合利用肿瘤基因组图谱(The Cancer Genome Atlas,TCGA)数据库和基因表达数据库(Gene Expression Omnibus,GEO)中的GSE130779和GSE85841数据集,分析PITX1基因在肺腺癌患者癌组织及癌旁组织中的表达水平,并利用实时荧光定量PCR法在40例肺腺癌患者组织中验证PITX1基因的表达水平。同时利用COX回归分析PITX1基因与肺腺癌患者的总生存期(overall survival,OS)和无复发生存期(recurrence-free survival,RFS)之间的相关性,进而分析其表达水平与肺腺癌患者临床病理特征之间的相关性。结果:基于TCGA和GEO数据库分析结果显示PITX1基因在肺腺癌组织中显著高表达(P<0.01),实时荧光定量PCR结果显示PITX1基因在肺腺癌组织的表达量显著高于癌旁正常组织(P<0.001),其相对表达量分别为1.064±0.077和0.641±0.044。COX回归分析显示PITX1基因的表达水平与肺腺癌患者的TNM分期、淋巴结转移状态和肿瘤大小显著相关(均P<0.05)。同时,上调PITX1的表达水平与肺腺癌患者的OS和RFS呈负相关。结论:PITX1基因在肺腺癌组织中显著高表达,且与肺腺癌患者的预后显著相关。     

Prognostic value of miR-26a and HMGA1 in urothelial bladder cancer

BackgroundMicroRNA-26a (miR-26a) functions as a tumor suppressor by regulating its direct target gene high mobility group AT-hook 1 (HMGA1). This study was aimed to investigate the associations of differential expression of miR-26a and HMGA1 with tumor progression and prognosis in urothelial bladder cancer (UBC) patients.Materials and methodsOne hundred and twenty-six UBC patients were selected and quantitative real-time PCR was performed to detect the expression of miR-26a and HMGA1 mRNA in the respective tumors.ResultsOur data showed the decreased expression of miR-26a and the increased expression of HMGA1 mRNA in UBC tissues compared with corresponding non-cancerous tissues (both P < 0.001). Then, the expression levels of miR-26a in UBC tissues were negatively correlated with those of HMGA1 mRNA significantly (r = –0.72, P < 0.001). In addition, UBC patients with combined miR-26a downregulation and HMGA1 upregulation (miR-26a-low/HMGA1-high) more frequently had advanced pathological stage (P < 0.001) and high tumor grade (P < 0.001). Moreover, miR-26a-low/HMGA1-high expression was associated with a significantly shortest disease-free survival (P < 0.001) and overall survival (P < 0.001) of all miR-26a/HMGA1 combined expression groups. Furthermore, multivariate analysis indicated that miR-26a/HMGA1 expression was an independent prognostic factor for both disease-free survival and overall survival (both P = 0.001) in UBC patients.ConclusionInteraction between miR-26a and its target gene HMGA1 may contribute to the malignant progression of human UBC. Tumors with miR-26a downregulation in combination with high expression of HMGA1 showed a worse prognosis than the other tumors. Combined detection of their expression might be particularly helpful for surveillance of disease progression and treatment stratification.     

LATS2 as a poor prognostic marker regulates non-small cell lung cancer invasion by modulating MMPs expression

Large tumor suppressor 2 (LATS2) plays significant roles in tumorigenesis and cancer progression. This study was aimed to analyze the correlation between LATS2 expression and clinicopathologic features and its prognostic significance in non-small cell lung cancer (NSCLC). LATS2 expression was examined in 73 NSCLC clinical specimens and 22 normal lung tissues using immunohistochemistry. Low levels of LATS2 protein were inversely associated with the T classification (P = 0.001), N classification (P = 0.005) and clinical stage (P = 0.001) in NSCLC patients. Patients with lower LATS2 expression had a significantly shorter overall survival than patients with high LATS2 expression. Multivariate analysis suggested that low expression of LATS2 was an independent prognostic indicator (P = 0.002) for the survival of patients with NSCLC. Furthermore, overexpression of LATS2 resulted in mobility inhibition in NSCLC cell lines A549 and H1299, and reduced protein level of matrix metalloproteinase-2 (MMP-2) and MMP-9. On the contrary, LATS2 siRNA treatment enhanced cell mobility and increased MMP-2 and MMP-9 protein expression level. In conclusion, low expression of LATS2 is a potential unfavorable prognostic factor and promoted cell invasion and migration in NSCLC.     

Can plasma DNA monitoring be employed in personalized chemotherapy for patients with advanced lung cancer?

Personalized chemotherapy is the ideal treatment usually chosen to help improve the survival chances of patients with advanced lung cancer. However, there is no short-term evaluation protocol for predicting the efficacy of the therapy. The aim of this study was to determine the value of using plasma DNA to monitor chemotherapeutic efficacy and to select most appropriate chemotherapeutic regimen for patients with advanced lung cancer. Eighty-eight lung cancer patients and 200 healthy controls were included in this study. Plasma DNA was extracted from plasma samples with internal controls by using the BILATEST DNA Kit. The quantity of plasma DNA was determined by using duplex real-time quantitative PCR. After first-line chemotherapy, plasma DNA levels of partial response patients were significantly different from those of stable disease patients or progressive disease patients, but with no statistical difference from healthy controls (P = 0.014, P < 0.001 and P = 0.418, respectively). Survival analysis showed a statistically better survival time in patients who had lower levels of plasma DNA after the third cycle chemotherapy (P = 0.031). In this study, the correlation of the kinetics of DNA concentrations with chemotherapeutic efficacy during the whole therapy was also observed. The quantification of plasma DNA is a sensitive indicator of chemotherapeutic efficacy in advanced lung cancer patients, and it can be useful in predicting response to therapy and guiding medication.     

Chronodisruption in lung cancer and possible therapeutic approaches

A customary temporal organization of physiological functions and biological processes is necessary to maintain body homeostasis and an altered body time structure may favour carcinogenesis. There is growing evidence that GH stimulates cancer growth, IGF1 may have a role in carcinogenesis and cancer promotion, GH-IGF1 axis, TRH, TSH, thyroxine, melatonin and cortisol modulate immune cell function and the immune system is often dysfunctional in patients with malignancies. The aim of our study was to evaluate GH-IGF1 axis, hypothalamus-pituitary-thyroid axis, melatonin, cortisol, lymphocyte subsets and IL2 in lung cancer patients. Peripheral blood samples were collected at 4-hour intervals in a 24-hour period from eleven healthy male subjects (age range 35-53 years) and nine male patients suffering from non-small cell lung cancer (age range 43-63 years). In each blood sample, lymphocyte subpopulations (CD3+, CD4+, CD8+, CD16+, CD20+, CD25+, HLA-DR+, γδTcR bearing cells) were analyzed and GH, IGF1, TRH, TSH, FT4, melatonin, cortisol and IL2 were measured. Circadian rhythmicity was evaluated and MESOR, amplitude and acrophase values were compared. In healthy subjects a significant circadian rhythm could be demonstrated with midday peaks for CD8+, CD16+, γδTCR expressing cells and cortisol, and peaks during the night for CD3+, CD4+, GH, TSH and melatonin. A borderline significant rhythm was also observed for CD20+, with a peak late in the evening. IGF1, TRH, FT4 and IL2 values did not show rhythmic variation. In cancer patients a significant circadian rhythm could be demonstrated with diurnal peak for CD16+ and peaks during the night for CD4+ and melatonin. GH, IGF1, TRH, TSH, FT4, cortisol and IL2 values did not show rhythmic variation. MESOR of CD8+ (P < 0.0001), CD20+ (P = 0.05), γδTCR expressing cells (P = 0.01), IGF1 (P < 0.001) and TSH (P = 0.032) was higher in healthy subjects, whereas MESOR of CD16+ (P < 0.0001), CD25+ (P = 0.001), GH (P < 0.001), TRH (P = 0.002), FT4 (P = 0.030), cortisol (P = 0.01) and IL2 (P = 0.02) was higher in cancer patients. Amplitude of circadian variation of γδTCR expressing cells (P = 0.01), TSH (P < 0.001) and cortisol (P = 0.01) was higher in healthy subjects, whereas amplitude of circadian variation of CD4+ was higher in cancer patients (P = 0.02). In conclusion, non-small cell lung cancer patients show severe alterations of periodic and quantitative characteristics of neuroendocrine and immune parameters with loss of circadian rhythmicity and internal desynchronization, leading to chronodisruption.     

Leptin and HER-2 are associated with gastric cancer progression and prognosis of patients

We conducted this study to evaluate the expression of leptin and its receptor, OB-Rb in gastric cancer and their relationship to clinicopathological features, VEGF and HER-2 expression, as well as the prognostic value. One hundred and ten gastric cancer specimens were detected for leptin, OB-Rb, VEGF and HER-2 by immunohistochemistry (IHC), and 96 specimens of normal gastric mucosa served as the control. The expression level of leptin, OB-Rb and HER-2 in gastric tissues were significantly higher than normal tissues (49.1% vs. 34.0%, 60.9% vs. 46.0%, 19.1% vs. 8.0%, P < 0.05). There was a correlation between the expression of leptin and HER-2, both of which were significantly associated with invasion depth, lymph node metastasis, AJCC stage and VEGF expression. However, there was no correlation between OB-Rb expression and all clinicopathological features. Cox regression analyses showed that age, tumor size, histological grade, serosa invasion, AJCC stage, chemotherapy, leptin and HER-2 overexpression were prognostic factors. The survival of patients with leptin positive expression was significantly poorer than those with negative expression (OS: 20.0 months vs. 32.5 months, FPS: 12.0 months vs. 18.0 months, P < 0.01). Leptin, rather than OB-Rb, played an important role in the progression and angiogenesis of gastric cancer, and was a prognostic factor for poor outcome.