Jinghao Liu

Circulating tumor DNA identified by targeted sequencing in advanced-stage non-small cell lung cancer patients

Non-small cell lung cancers (NSCLC) have unique mutation patterns, and some of these mutations may be used to predict prognosis or guide patient treatment. Mutation profiling before and during treatment often requires repeated tumor biopsies, which is not always possible. Recently, cell-free, circulating tumor DNA (ctDNA) isolated from blood plasma has been shown to contain genetic mutations representative of those found in the primary tumor tissue DNA (tDNA), and these samples can readily be obtained using non-invasive techniques. However, there are still no standardized methods to identify mutations in ctDNA. In the current study, we used a targeted sequencing approach with a semi-conductor based next-generation sequencing (NGS) platform to identify gene mutations in matched tDNA and ctDNA samples from 42 advanced-stage NSCLC patients from China. We identified driver mutations in matched tDNA and ctDNA in EGFR, KRAS, PIK3CA, and TP53, with an overall concordance of 76%. In conclusion, targeted sequencing of plasma ctDNA may be a feasible option for clinical monitoring of NSCLC in the near future.

Detection of circulating tumor DNA in patients with advanced non-small cell lung cancer

Circulating tumor DNA (ctDNA) isolated from plasma has great potential in identification of gene mutation in non-small cell lung cancers (NSCLC), which is a non-invasive technique and can avoid the inherent shortcomings of tissue biopsy. However the ability of NGS to detect gene mutation in plasma ctDNA has not been broadly explored. To assess the diagnostic ability of ctDNA for the total mutation profile, including single nucleotide variations (SNVs), insertions and deletions (indels) and gene rearrangements, we performed a targeted DNA sequencing approach to screen NSCLC related driver gene mutations in both tissue biopsies and matched blood plasma samples from 39 advanced NSCLC patients from China. The sensitivity of EGFR, KRAS, PIK3CA mutations and gene rearrangements detected in plasma ctDNA was 70.6%, 75%, 50% and 60%, respectively and the overall concordance of gene mutations between tissue DNA and plasma ctDNA was 78.21%. Our data provide evidence that ctDNA in plasma is likely to become an alternative source for cancer-related mutations profiling in advanced NSCLC patients and targeted sequencing of ctDNA offers a promising perspective on precise diagnostics and may serve as a feasible option for clinical monitoring of NSCLC patients.

MiR-146b-5p functions as a suppressor miRNA and prognosis predictor in non-small cell lung cancer

Non-small cell lung cancer (NSCLC) is the leading cause of cancer-related death worldwide. However, science has not yet been able to substantially improve the prognosis of lung cancer patients. Accumulating evidence suggests that microRNAs (miRNAs) are key players in the regulation of tumor development and metastasis. Expression of six miRNAs previously shown to play roles in tumor development (miR-146b-5p, miR-128b, miR-21, miR-221, miR-34a, and Let-7a) in other tumor types was examined using real-time RT-PCR in 78 specimens of NSCLC. The results revealed that patients with low expression of miR-146b-5p had significant shorter median and mean survival time than those with high miR-146b-5p expression (33.00 and 30.44 months versus 42.0 and 36.90 months, respectively; log-rank test P=0.048), thus low miR-146b-5p expression level was associated with poor prognosis in NSCLC patients. Univariate Cox hazard regression analysis demonstrated that miR-146b-5p expression levels tended to be a significant prognostic indicator of NSCLC (adjusted hazard ratio=0.482, 95% CI: 1.409- 29.593, P=0.016). Multivariate Cox proportional hazard regression analysis showed that miR-146b-5p expression levels were an independent prognostic factor for NSCLC patients (hazard ratio=0.259, 95% CI: 0.083-0.809, P=0.020). Furthermore, the effects of miR-146b-5p and miR-146b-3p on NSCLC cell growth and invasion in vitro were investigated. Our findings demonstrate that ectopic expression of miR-146b-5p suppressed cell proliferation, clonogenicity, migration/ invasion and also induced G1 arrest in vitro, but did not induce cell apoptosis; whereas enforced expression of miR-146b-3p did not have a significant effect on cell growth and metastasis. Further experiments indicated that miR-146b-5p could reduce mRNA levels of MMP16 and TRAF6 in vitro and was negatively related to the expression of TRAF6 in human NSCLC tissues. In a mouse model, Ago-miR-146b-5p could significantly inhibit the growth of lung cancer xenografts in nude mice. In conclusion, our findings demonstrate that miR-146b-5p functions as a suppressor miRNA and prognosis predictor in NSCLC.

Thyroid transcription factor-1 expression is significantly associated with mutations in exon 21 of the epidermal growth factor receptor gene in Chinese patients with lung adenocarcinoma.

Objective The aim of this retrospective study was to investigate the relationship between thyroid transcription factor-1 (TTF-1) expression and epidermal growth factor receptor (EGFR) gene mutations in lung adenocarcinomas of Chinese patients. Methods There were 200 lung adenocarcinoma patients who were enrolled in this study. Tumor specimens of these patients were investigated for TTF-1 expression and mutations in EGFR using immunohistochemistry and a liquid chip platform for DNA analysis of slides with sections of formalin-fixed, paraffin-embedded specimens. Results The rates of TTF-1 expression and EGFR mutations were 81.5% and 45.5%, respectively, in the lung adenocarcinoma specimens of the recruited patients. Among female nonsmokers (n=72), 93.1% of specimens were positive for TTF-1 expression, and 63.9% had EGFR mutations. Of 89 patients with EGFR mutations, 83 (50.9%) specimens were simultaneously positive for TTF-1 expression. Kaplan–Meier analysis of all patient specimens found that postoperative survival time was not significantly associated with TTF-1 expression and the presence of EGFR mutations. However, patients with disease stages III–IV whose tumors were positive for TTF-1 expression and EGFR mutations had better postoperative survival than similar patients whose tumors were negative for TTF-1 expression and EGFR mutations. Conclusion Our study showed a significant association between TTF-1 positivity and the presence of EGFR mutations (exon 21) in the Chinese lung adenocarcinoma patients. We further identify that patients with disease stages III–IV who were positive for TTF-1 expression and EGFR mutations had a better postoperative survival than those patients who were negative for TTF-1 expression and EGFR mutations. Therefore, TTF-1 might be a potential prognostic biomarker for stages III–IV lung adenocarcinoma patients. In clinical practice, TTF-1 expression may be a marker for planning therapy for certain patients with lung adenocarcinoma, especially for selection of EGFR tyrosine kinase inhibitors.

MiR-182 inhibits the epithelial to mesenchymal transition and metastasis of lung cancer cells by targeting the Met gene

The microRNA miR‐182, belonging to the miR‐183 family, is one of the most frequently studied cancer‐related oncogenic miRNAs that is dysregulated in various cancer tissues, and it plays a crucial role in tumorigenesis and tumor progression. Studies revealed that miR‐182 might function as an oncogenic or tumor suppressor miRNA in different tissues. However, the role of miR‐182 in the development of lung cancer remains largely unknown. miR‐182 expression in tumor samples from 58 patients, normal lung tissue samples, and lung cancer cell lines were evaluated by qRT‐PCR. Survival curves were analyzed using the Kaplan‐Meier method and compared with a log‐rank test. Our study demonstrated that miR‐182 is frequently downregulated in metastatic NSCLC cells compared with primary tumor tissues. Over‐expression of miR‐182 significantly inhibited the migration and invasion of lung cancer cells and promoted the expression of the epithelial marker (E‐cadherin) in addition to reducing the levels of Snail in lung cancer cells. Further studies demonstrated that miR‐182 negatively regulated Met via direct binding to the Met 3′‐untranslated region (3′‐UTR). Furthermore, we found that miR‐182 suppressed the phosphorylation of AKT and the nuclear accumulation of Snail, a transcription factor that promotes the epidermal to mesenchymal transition (EMT). Moreover, miR‐182 could repress cell migration, invasion, and EMT of lung cancer cells induced by hepatocyte growth factor (HGF). miR‐182 might suppress the EMT and metastasis via inactivation of Met/AKT/Snail in non‐small cell lung cancer (NSCLC) cells, which implicates miR‐182 may be useful as a new therapeutic target in NSCLC.

Influence of Thoracoscopic Surgery on Inflammatory Reaction of the Body for Early Peripheral Lung Cancer Patients

BACKGROUND AND OBJECTIVE It has been proven that video assited thoracoscopic surgery (VATS) achieved the same survival rates compared with traditional open chest operation in the treatment of early stage of lung cancer. but it is unclear if there is any difference of body inflammatory reaction between the two operation. The aim of this study is to investigate the changes of inflammatory state of thoracoscopic radical lobectomy in early peripheral lung cancer patients. METHODS Senventy-one early peripheral lung cancer patients who have underwent radical lobectomy were divided into two groups based on the different operation method. The VATS group was treated by thoracoscopic lobectomy. The thoracotomy group was treated by traditional thoracotomy. Then the level of serum C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6) and IL-10 at 1-day before operation and 3-day, 7-day postoperation were measured and compared between the two groups. RESULTS No significant difference was found in the level of serum CRP, TNF-α, IL-6 and IL-10 before operation. Compared to the thoracotomy group, the level of serum CRP, TNF-α, IL-6 and IL-10 in the VATS group were significantly lower after operation. CONCLUSIONS Compared with thoracotomy lobectomy, thoracoscopic lobectomy for early peripheral lung cancer patients is associated with lower inflammatory responses .

Clinicopathological characteristics and molecular analysis of primary pulmonary mucoepidermoid carcinoma: Case report and literature review

Primary pulmonary mucoepidermoid carcinoma (PMEC) is extremely rare. Herein, we report a case of a 71‐year‐old male patient with high‐grade PMEC involving the right upper lobe that was successfully resected via lobectomy. As a result of invasion into the pleural and paratracheal lymph nodes, four cycles of adjuvant chemotherapy with paclitaxel and carboplatin were administered. There were no signs of relapse during 10 months of follow‐up. Furthermore, we reviewed the literature and summarized the surgical approaches, prognostic factors, and underlying genetic mechanisms of PMEC, which will benefit clinical treatment.

Role of artemin in non-small cell lung cancer: Function of artemin in lung cancer

In this study, we investigated the role of artemin, a member of the glial cell‐derived neurotrophic factor of ligands, in the malignant phenotype of lung cancer.

Molecular features of giant-cell carcinoma of the lung: a case report and literature review

Giant-cell carcinoma of the lung (GCCL) is a rare histological form of poorly differentiated non-small-cell lung cancer, which is classified as a subtype of pulmonary sarcomatoid carcinomas. In this case report, we describe the case of a 50-year-old Chinese male who presented with a pulmonary nodule in the right upper lobe of his lung. After thoracoscopic lobectomy, a histopathologic diagnosis of GCCL was made. He did well postoperatively, showing no local recurrence or distal disease in a 7-year follow-up period. Furthermore, for this case, we also analyzed 295 tumor-related driver genes with high-throughput sequencing technology. We found that treatment using MEK inhibitor, CDK 4/6 inhibitor, and TP53 inhibitor may provide a new therapeutic direction for GCCL. Therefore, complete tumor excision is the best choice of treatment strategy at the early stage of GCCL and gene target therapy may be a new therapeutic option for this disease.

Clinicopathological and genetic characteristics of pulmonary large cell carcinoma under 2015 WHO classification: a pilot study

Pulmonary large cell carcinoma (LCC) was re-defined under the 2015 WHO classification criteria. However, the clinicopathological features and genetic mutation statuses of Chinese LCC patients based on the new classification have rarely been investigated. Twenty-four Chinese surgically resected LCC patients previously diagnosed under the 2004 WHO criteria were re-classified under the 2015 WHO criteria. Genetic analysis was performed using next-generation sequencing of 46 cancer-related genes. The correlation of clinicopathological and genetic data was further analyzed. Eight patients were re-defined as LCCs, and 16 patients were defined as non-LCCs under the refined criteria. All LCC patients were male, and 7 patients were smokers. No significant differences in age, gender, smoking status, primary site, TNM staging and overall survival were observed between the LCC and non-LCC patients under the refined criteria. Four of the 8 LCC patients presented TP53 mutations, and no somatic mutations were detected in the other 4 LCCs under the refined criteria. For the 16 non-LCCs, not only TP53 and KRAS but also EGFR, KIT, PIK3CA, PTEN, IDH1, APC, ATM and BRAF mutations were also observed. In addition, LCCs without TP53 mutations did not present any gene mutations under the 2004 or 2015 WHO criteria. Importantly, the patients with TP53 mutation exhibited a trend with a worse survival outcome at the time of follow-up. The new WHO diagnosis criteria have superior performance in precise molecular classification for LCC patients.