Guangqiang Zhao

Down‐regulated SOX4 Expression Suppresses Cell Proliferation, Metastasis and Induces Apoptosis in Xuanwei Female Lung Cancer Patients

The transcription factor SOX4 has functional importance in foetal lung maturation and tumorigenesis in a number of cancers. However, its biological functions in the progression of lung tumorigenesis remain unclear. In this study, we found that the expression levels of SOX4 mRNA and protein were significantly higher in Xuanwei female lung cancer tissues than in benign lung lesions. The patients with high expression of the SOX4 protein had a higher pathological grade, lymph node (LN) metastasis, poor tumor differentiation and worse prognosis than those patients with low expression of SOX4. Knockdown of the SOX4 gene in the Xuanwei female lung cancer cell line XWLC‐05 resulted in apoptotic morphological changes, decreased cell proliferation, invasion and migration. Furthermore, knockdown of the SOX4 gene resulted in obvious sub‐G1 peaks and induction of apoptosis through upregulation of caspase‐3 expression, while in cells treated with a caspase‐3 inhibitor, apoptosis induced by silencing SOX4 expression was inhibited. In vivo analysis in nude mice further confirmed that knockdown of SOX4 suppressed tumor growth. In conclusion, SOX4 appears to be an important tumor suppressor gene in the regulation of Xuanwei female lung cancer cell proliferation, apoptosis and metastases, and it may be a potential target for effective lung cancer therapy. J. Cell. Biochem. 116: 1007–1018, 2015.

The clinical use of circulating microRNAs as non-invasive diagnostic biomarkers for lung cancers

Many studies have investigated the diagnostic role of circulating microRNAs (miRNAs) in patients with lung cancer; however, the results still remain inconclusive. An updated system review and meta-analysis was necessary to give a comprehensive evaluation of diagnostic role of circulating miRNAs in lung cancer. Eligible studies were searched in electronical databases. The sensitivity and specificity were used to plot the summary receiver operator characteristic (SROC) curve and calculate the area under the curve (AUC). The between-study heterogeneity was evaluated by Q test and I2 statistics. Subgroup analyses and meta-regression were further performed to explore the potential sources of heterogeneity. A total of 134 studies from 65 articles (6,919 patients with lung cancer and 7,064 controls) were included for analysis. Overall analysis showed that circulating miRNAs had a good diagnostic performance in lung cancers, with a sensitivity of 0.83, a specificity of 0.84, and an AUC of 0.90. Subgroup analysis suggested that combined miRNAs and Caucasian populations may yield relatively higher diagnostic performance. In addition, we found serum might serve as an ideal material to detecting miRNA as good diagnostic performance. We also found the diagnostic role of miRNAs in early stage lung cancer was still relatively high (the sensitivity, specificity and an AUC of stage I/II was 0.81, 0.82 and 0.88; and for stage I, it was 0.80, 0.81, and 0.88). We also identified a panel of miRNAs such as miR-21-5p, miR-223-3p, miR-155-5p and miR-126-3p might serve as potential biomarkers for lung cancer. As a result, circulating miRNAs, particularly the combination of multiple miRNAs, may serve as promising biomarkers for the diagnosis of lung cancer.

Epidermal growth factor receptor (EGFR) mutations in non-small cell lung cancer (NSCLC) of Yunnan in southwestern China

To investigate the Epidermal Growth Factor Receptor (EGFR) mutation status in non-small cell lung cancer (NSCLC) in Yunnan province in southwestern China, we detected EGFR mutation by Amplification Refractory Mutation System (ARMS) polymerase chain reaction (PCR) using DNA samples from 447 pathologically confirmed NSCLC specimens (175 tissue, 256 plasma and 16 cytologic samples). The relationship between EGFR mutations and demographic and clinical factors were further explored. Subgroup analyses according to sample type (tissue and plasma) and histological type (adenocarcinoma) were done. We found the mutation rate was 34.9% in overall patients (42.3%, 29.7%, and 37.5% for tissue, plasma, and cytologic samples respectively). We found female (p < 0.0001), no smoking (p = 0.001), adenocarcinoma (p < 0.0001), and tissue specimen (p = 0.026) were associated with higher EGFR mutation rate. The most common mutations were exon 19 deletions (40%) and L858R point (30%) mutation. Interestingly, NSCLC patients from Xuanwei harbored a strikingly divergent mutational pattern for EGFR when compared with non-Xuanwei patients (higher G719X, G719X+S768I mutations, but lower 19 deletion and L858R mutations). Generally, EGFR mutation rate and pattern in Yunnan province was in accord with other Asian populations. However, Xuanwei subgroup showed strikingly divergent EGFR mutation spectrum from other general population. Our analysis also indicated that cftDNA analysis for EGFR mutations detection was feasibility for the patients lacking sufficient tissue for molecular analyses.

Tree shrew as a new animal model for the study of lung cancer

Animal models play a key role in identifying treatments for various types of cancer, including lung cancer. The aim of the present study was to develop a new animal model for lung cancer induction using tree shrews from the Yunnan region in China. Tree shrews are suitable for a full simulation of human disease because their structure, function and metabolism are adequately close to human. This animal may offer a new experimental animal model to be used in the study of lung cancer. In the present study, 80 healthy tree shrews were distributed in experimental and control groups. Animals in the experimental group received different concentrations of iodized oil suspension of 3-methylcholanthrene (3-MC) and diethylnitrosamine (DEN) while animals in the control groups received saline or lipiodol solvent via endotracheal instillation. In the 3rd, 5th, 7th, 9th and 11th weeks the body weights of the animals were measured and chest X-ray examinations were conducted. Pathological studies on the lung tissues were also performed and the pathological changes occurring in bronchial epithelium in all the groups were examined. Animals in the experimental group gradually lost their body weight. For tree shrews in the blank control and solvent control groups the survival rates were 100 and 80%, respectively while the survival rate for the experimental group was 0%. Results from the chest X-ray conducted on animals in the blank control and solvent control groups revealed no obvious abnormalities while in the experimental group high-density shadow spots within the perfusion sites were observed. Pathological studies performed on these high-density areas confirmed changes in the bronchial epithelium. In the experimental groups we also detected bronchial epithelial atypical hyperplasia, and apparent changes in carcinoma in situ. In conclusion, lung cancer was successfully induced in tree shrews by a one-time endotracheal introduction of iodized oil suspension of 3-MC and DEN.

Subcellular distribution and genotoxicity of silica nanoparticles in human bronchial epithelial cells

BACKGROUND Silicon nanoparticles are widely used in daily life. Therefore, they attract increased attention because of their potential biotoxicity to the lungs when inhaled. The aims of this study are to explore the organism distribution and genotoxicity of silica nanoparticles in human bronchial epithelial cells (BEAS-2B). METHODS The biodistribution of silica with different particle sizes in human bronchial epithelial cells was observed by transmission electron microscopy (TEM). DNA damage was detected by single-cell gel electrophoresis (comet assay). RESULTS TEM revealed that SiO₂ nanoparticles with different sizes can be uptaken by cells and be localized in the cytoplasm and the nucleus. Compared with micro-silica, nano-silica in BEAS-2B cells can inflict more severe DNA damage (P<0.05). CONCLUSIONS The particle size of silica nanoparticles can be used to determine their distribution in biological cells. Compared with micro-silica, nano-silica has higher genotoxicity.

Research on fast track surgery application in lung cancer surgery

背景与目的 快速康复外科(fast track surgery, FTS)通过减少手术病人生理和心理的创伤应激, 从而达到病人快速康复的目的。本研究旨在探讨FTS技术在肺癌手术中应用的可行性。 方法 选取80例适合单叶肺叶切除手术治疗的肺癌患者, 随机分为实验组及对照组, 实验组接受FTS方案治疗、对照组接受传统方法治疗。比较两组术后不同时间的疼痛程度、肺不张发生率、胸腔积液发生率、术后住院时间及住院总费用。 结果 FTS方案组:术后1 h、6 h、12 h、24 h、48 h疼痛VAS评分均明显低于传统治疗组; 肺不张发生率为10.53%, 胸腔积液发生率为26.31%, 术后住院时间(4±1)d, 住院总费用(1.56 ±0.76)万元。对照组:肺不张发生率为33.33%, 胸腔积液发生率为22.22%, 术后住院时间(9±1)d, 总费用(2.36 ±0.54)万元。两组比较:术后疼痛程度(视觉模拟评分)明显低于对照组, 差异有统计学意义(P < 0.01);肺不张发生率(P=0.035), 住院时间(P=0.021)及住院总费用(P=0.024)的组间差异均有统计学意义; 胸腔积液发生率的组间差异无统计学意义(P=0.223)。 结论 FTS方案的应用可有效促进肺癌患者术后的康复, 减少术后并发症发生, 缩短住院时间, 降低住院费用。BACKGROUND AND OBJECTIVE Fast track surgery (FTS) is a systematical method to accelerate the recovery of surgical patients by reducing the physical and mental trauma stress of them. The research is to investigate the feasibility of FTS application in lung cancer surgery. METHODS A total of 80 cases of lung cancer patients with single leaf lobotomy resection were randomized into two groups. While the experimental group was treated with the conception of FTS, and the control group was treated with the traditional methods. The incident rate of post-operation pain degrees, telecasts, pleural effusion, the post-operation time stay in hospital time and the total cost during hospitalization in two groups were compared respectively. RESULTS In FTS group: the VAS score of post-operation pain at 1 h, 6 h, 12 h, 24 h and 48 h all significantly decreased compared to the traditional therapy group. The incidence rate of telecast was 10.53%. The incidence rate of pleural effusion was 26.31%. The length of stay after operation was (4 +/- 1) d and the total cost was RMB 15 600 +/- 7 600. In the control group, the above values were 77.78%, 33.33%, 22.22%, (9 +/- 1) d, RMB 23 600 +/- 5 400, respectively. The post operation pain (VAS method) of FTS group was remarkablely below the control group. There has significant difference of the incident rate of telecasts, stay time in hospital and the total cast in two groups (P < 0.05). No significant difference was observed in the incident rate ofpleural effusion. CONCLUSION The new methods of FTS can apparently accelerates recovery after lung cancer resection, reduces complications, shorten timestay in hospital and cut down the total cost.

Temporal trend of mortality from major cancers in Xuanwei, China

Although a number of studies have examined the etiology of lung cancer in Xuanwei County, China, other types of cancer in this county have not been reported systematically. This study aimed to investigate the temporal trend of eight major cancers in Xuanwei County using data from three mortality surveys (1973–1975, 1990–1992, and 2004–2005). The Chinese population in 1990 was used as a standard population to calculate agestandardized mortality rates. Cancers of lung, liver, breast, brain, esophagus, leukemia, rectum, and stomach were identified as the leading cancers in this county in terms of mortality rate. During the three time periods, lung cancer remained as the most common type of cancer. The mortality rates for all other types of cancer were lower than those of the national average, but an increasing trend was observed for all the cancers, particularly from 1990–1992 to 2004–2005. The temporal trend could be partly explained by changes in risk factors, but it also may be due to the improvement in cancer diagnosis and screening. Further epidemiological studies are warranted to systematically examine the underlying reasons for the temporal trend of the major cancers in Xuanwei County.

Effects of Benzoapyrene on migration and invasion of lung cancer cells functioning by TNF-α

In this study, we attempted to find out the underlying mechanism of Benzoapyrene and metastasis of lung cancer cells. We also did experiments to testify the connection between BaP and its potential target, TNF‐α. Cell median lethal dose (IC50) of both cells was measured by crystal violet method. Quantitative real‐time reverse transcription PCR (qRT‐PCR) and Western blot were employed to detect the expression of TNF‐α. Wound healing assay and transwell assay were utilized to testify the impacts of BaP and TNF‐α on the metastasis of lung cancer cells. Cell death rate was elevated with the increase of BaP concentration. BaP increased the number of metastatic cells of lung cancer. The expressions of TNF‐α pathway‐associated protein (TNF‐α, NF‐kB [P65], Caspase3, and Caspase8) were enhanced by overexpressed BaP. TNF‐α shRNA suppressed the positive effects of BaP on migration and invasion of lung cancer cells. Our study validated the positive effects of BaP on the metastasis of lung cancer cells. We also revealed the instrumental role of TNF‐α in helping the development of lung cancer cells induced by BaP.

Expression of PANDA, LincRNA-p21, PUMA in lung tissues of lung cancer patients in the Xuanwei and non-Xuanwei areas of Yunnan Province

Aim: To study the expression of PANDA, LincRNA-p21, and PUMA in lung tissue of patients with lung cancer from Xuanwei of Yunnan Province. Methods: Forty-five cases of lung cancer patients from Xuanwei and 42 lung cancer cases from non-Xuanwei were enrolled. Extraction of RNA was done using the Trizol kit. Real-time fluorescence quantitative PCR assay was done to obtain the relative expression. Results: Expressions of PANDA, LincRNA-p21, and PUMA in male and female patients or in squamous cell carcinoma and adenocarcinoma were not significantly different (P > 0.05). However, expression of LincRNA-p21 in Xuanwei patients was higher than non-Xuanwei patients (P < 0.05). Expression of PUMA in tumor tissue was lower than that in normal lung tissue (P < 0.05), and in Xuanwei patients was lower than non-Xuanwei patients (P < 0.05). In patients from non-Xuanwei regions, expression of LincRNA-p21 in patients with smoking index > 400 was higher than in those < 400 and non-smokers. Conclusion: Expressions of PANDA, LincRNA-p21, and PUMA in lung tissues have no gender differences or tissue specificity. High expression of LincRNA-p21 in Xuanwei patients may have relationship with cell damage caused by coal burning pollution in Xuanwei.

Association of Inorganics Accumulation with the Activation of NF-κB Signaling Pathway and the iNOS Expression of Lung Tissue in Xuanwei Lung Cancer Patients

BACKGROUND AND OBJECTIVE Indoor air pollution induces asthma, leads to chronic obstructive pulmonary disease, and may promote lung cancer. Our previous studies found that the accumulation of inorganic particulate matter that is due to indoor air pollution can lead to damage to alveolar cells and activation of signaling pathway, and ultimately provoke tumorigenesis. The aim of this study is to explore the accumulation of inorganics and activation of nuclear factor κB (NF-κB)-inducible nitric oxide synthase (iNOS) signaling pathway of lung tissue in Xuanwei lung cancer patients. METHODS From December 2013 to November 2014, 48 cases Xuanwei patients with lung cancer who underwent surgical treatment from the Third Affiliated Hospital of Kunming Medical University were enrolled in this study and compared with lung cancer patients from other regions. The ultrastructure of postoperative specimens was observed by transmission electron microscopy (TEM) to explore the occurrence of inorganic particles. Serum cytokines were analyzed. Then, the expression levels of NF-κB-p65 protein and iNOS protein in postoperative specimens was explored by immunohistochemistry and Western blot. Finally, 8-OHdG accumulation in lung cancer tissues and urine was measured. RESULTS A large number of nanoscale inorganics were observed in alveolar type II cells and macrophages located in adjacent tissues of lung cancer with Xuanwei patients. Silicon (Si) content was found in inorganic elemental analysis. The serum interleukin (IL)-1β levels (31.50 ± 19.16) pg/mL of Xuanwei lung-cancer patients were remarkably higher than those from other regions (11.33 ± 6.94) pg/mL (P<0.01), with statistically significant difference. The pathological tissues of Xuanwei lung-cancer patients express NF-κB-p65, and iNOS expression were significantly higher than those of patients from non-Xuanwei regions. No significant difference was found between cancerous and normal adjacent tissues. Xuanwei lung-cancer tissues and urine 8-OHdG level (40.124 ± 8.597) ng/mgCr were significantly higher than those of patients from other regions (25.673 ± 7.986) ng/mg Cr (P<0.05), with statistically significant difference. CONCLUSIONS The accumulation of inorganics and the activation of NF-κB-iNOS signaling pathway may contribute to Xuanwei lung cancer.
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