Function and mechanism exploration of zinc finger protein 64 in lung adenocarcinoma cell growth and metastasis.

This paper aims to discover the effect of Zinc Finger Protein 64 (ZFP64) and Notch pathway on lung adenocarcinoma cell. ZFP64 expression in cancer tissue and overall survival analysis was identified by TCGA-LUAD. ZFP64 expressions in tumor tissue (n = 30) and adjacent tissue (n = 30), and in human nontumorigenic bronchial epithelial cell line BEAS-2B and human lung adenocarcinoma cell lines (H23, H1975, H2228, and H2085) were measured via quantitative real-time polymerase chain reaction (qRT-PCR). H1975 cell viability, cell cycle progression, and migration after transfection or under Notch inhibitor MK-0752 treatment were detected through MTT assay, flow cytometer, and wound healing assay, respectively. Expressions of notch intracellular domain (NICD) and hairy and enhancer of split 1 (Hes-1) in H1975 cell were determined by western blot. Epithelial-mesenchymal transition (EMT)-related proteins (E-Cadherin and Vimentin) expressions were identified through qRT-PCR and western blot. ZFP64 expression in lung adenocarcinoma tissue and lung adenocarcinoma cell lines was higher and related to poor prognosis. After transfection, H1975 cell viability, migration, and expressions of Vimentin, NICD and Hes-1 were upregulated yet cell percentage in G0/G1 phase, E-cadherin expression was downregulated by overexpressed ZFP64. However, Notch inhibitor MK-0752 inhibited the effects of overexpressed ZFP64 on H1975 cell viability, cell cycle, migration, EMT progress, and Notch pathway activation. Overexpressed ZFP64 promoted the development of lung adenocarcinoma cells by activating Notch pathway.

TBL1XR1 is involved in c-Met-mediated tumorigenesis of human nonsmall cell lung cancer.

Nonsmall cell lung carcinoma (NSCLC) contributes to the highest number of cancer deaths globally. Metastases and chemoresistance are two major confounders to the treatment efficacy in NSCLC. Transducin (ß)-like 1 X-linked receptor 1 (TBL1XR1) has been associated with high rates of metastases in breast, gastric, and stomach cancers. However, the role of TBL1XR1 in lung cancers remains underexplored. We selected matched and cancerous lung tissues to establish the upregulation of TBL1XR1. Using in vitro assays, we assessed the influence of TBL1XR1 on various cancer phenotypes, namely cell proliferation, chemoresistance, invasion, and metastases in a CRISPR-Cas9-mediated knock out model (A549 cells), and H460 cell lines overexpressing TBL1XR1. We found that TBL1XR1 is overexpressed in NSCLC tissue and patient sera in comparison to paired adjacent normal tissue. Overexpression of TBL1XR1 in NSCLC cell lines mediates cell survival, proliferation, and metastases. TBL1XR1 was found to regulate MEK and Akt pathways through their master regulator c-Met. We observed that activation of c-Met is downregulated in the absence of TBL1XR1. Our study strengthens the contention that TBL1XR1 is a biomarker for prognosis of NSCLC. It may also be considered as an adjunct or core therapeutic target to overcome cisplatin resistance in lung cancers.

LncRNA CCAT2 promotes tumorigenesis by over-expressed Pokemon in non-small cell lung cancer.

BACKGROUND: Non-small cell lung cancer (NSCLC) remains one of the most important death-related diseases, with poor effective diagnosis and less therapeutic biomarkers. LncRNA colon cancer-associated transcript 2 (CCAT2) was identified as an oncogenic lncRNA and over-expressed in many tumor cells. The aims of this study were to detect the correlation between CCAT2 and its regulatory genes and then explore the potential mechanism between them in NSCLC. METHODS: In this study, qRT-PCR was used to detect CCAT2, Pokemon and p21 expression. Western-blot was used to detect protein levels of Pokemon and p21. CCK-8 assay and Transwell chambers were used to assess cell viability and invasion. RESULTS: CCAT2 and Pokemon were over-expressed in NSCLC tissue and cells. In NSCLC cells, CCAT2 knockdown significantly decreased cell viability and invasion as well as Pokemon expression, but increased the expression of p21; then CCAT2 overexpression revealed an opposite result. In addition, over-expressed Pokemon reversed the results that induced by si-CCAT2, while down-regulation of Pokemon significantly reversed the results that induced by CCAT2 overexpression. CONCLUSION: The results indicated that CCAT2 promotes tumorigenesis by over-expression of Pokemon, and the potential mechanism might relate to the Pokemon related gene p21.

Pre-emptive epidural analgesia improves post-operative pain and immune function in patients undergoing thoracotomy.

BACKGROUND: Pre-emptive analgesia may relieve post-operative pain. However, its effects on immune function of patients undergoing thoracotomy are still unclear. Therefore, we investigated effects of pre-emptive epidural analgesia on post-operative pain and immune function in patients undergoing thoracotomy. METHODS: Ninety patients undergoing thoracotomy were randomized equally into three groups. Group A (control) only received patient-controlled epidural analgesia (PCEA). Group B (routine) was given 0.125% ropivacaine 6 mL 30 min after surgery, then PCEA. Group C (pre-emptive) received 0.125% ropivacaine 6 mL 30 min before skin incision and every 60 min during surgery, then PCEA. Visual analogue scale scores and cytokine levels were the main outcomes. Secondary outcomes included analgesic demands, side effects and hospital stays. RESULTS: Post-operative visual analogue scale scores were decreased in group C compared with group B, and that of group B were lower than in group A (P < 0.05). Compared with group B, protein and mRNA expressions of tumour necrosis factor-α, interleukin (IL)-6 and IL-8 were decreased, and that of IL-4 were increased in group C (P = 0.002∼0.049). Both analgesic demands and the incidences of nausea/vomiting and hypotension were lower in group C compared with group B (P = 0.01∼0.037). Hospital stays were shorter in group C compared with group B (P = 0.045). CONCLUSIONS: These findings suggest that pre-emptive epidural analgesia can produce better analgesia effects, with less analgesic demands and side effects, and attenuate the surgery-induced immune alterations, and improve the post-operative recovery in patients undergoing thoracotomy.

[Effect of cisplatin on the expression of Pokemon gene: experiment with different human lung cancer cells].

OBJECTIVE: To investigate the correlation between Pokemon gene and cisplatin mechanism. METHODS: Human lung adenocarcinoma cells of the lines A549 and AGZY83-a, human lung squamous carcinoma cells of the line HE-99, and human giant cell lung cancer cells of the line 95D were cultured and cisplatin was added into the medium. Other lung cancer cells of the above mentioned lines were cultured in the medium without cisplatin and were used as control groups. RT-PCR and Western blotting were used to detect the mRNA and protein expression of Pokemon. RESULTS: Pokemon mRNA and protein were expressed highly in all the 4 cell lines. The Pokemon gene expression did not changed significantly after cisplatin treatment groups. There were not significant differences in the mRNA and protein expression of Pokemon among the 4 experiment groups and the control groups (all P > 0.05). CONCLUSION: Cisplatin has no effect on the Pokemon gene expression of the human lung cancer cells.

[Suppression of RASSF1A gene on human esophageal carcinoma cells: experiments in vitro and in vivo].

OBJECTIVE: To investigate the effect of Ras association domain family 1A (RASSF1A) gene, a new tumor suppressor gene (TSG), on tumorigenesis of human esophageal carcinoma cells. METHODS: pcDNA3.1 (+)-RASSF1A, a plasmid containing RASSF1A gene, and the blank plasmid pcDNA3.1 (+) were transfected into human esophageal carcinoma cells of the line EC9706. The expression of RASSF 1A protein was examined by Western blotting. The changes of cell cycle of stably-transfected cells were determined by flow cytometry (FCM), and the cellular proliferation was analyzed by MTT assay. Fifteen nude mice were randomly divided into 3 groups to be inoculated subcutaneously with EC9706 cells transfected with pcDNA3.1 (+)-RASSF1A, EC9706 cells transfected with pcDNA3.1 (+), and untransfected EC9706 cells respectively. Other 5 nude mice were used as controls. Four weeks later, the mice were killed to take out the carcinoma tissues. FCM was used to analyze the cell cycle. RESULTS: Western blotting showed that RASSF1A protein was expressed highly in the stably transfected cells. The cell viability and growing speed were decreased obviously in the cells expressing of RASSF1A (both P < 0.01); FCM showed that the proportion of cells at the G(1) phase of the EC9706 cells expressing RASSF1A was significantly higher than those in the blank plasmid group and untransfected group (both P < 0.01). The size of the EC9706 cells obtained from the nude mice inoculated with the EC9706 cells transfected with pcDNA3.1 (+)-RASSF1A was significantly smaller than those of the pcDNA3.1 (+) group and blank plasmid group (both P < 0.05). CONCLUSION: Expression of exogenous RASSF1A inhibits the progression of human esophageal carcinoma cells in vitro and in vivo. As a tumor suppressor gene, it plays an important role in origination, progression and metastasis of esophageal carcinoma.

[Expression and clinical significance of Pokemon in non-small cell lung cancer].

BACKGROUND: Proto-oncogene Pokemon is the special transcription inhibitor of ARF,which can regulate cell growth and differentiation by ARF-P53 path.It may be the important monitoring target of tumor because of being upstream region of many tumor suppressor genes and proto-oncogenes.The aim of this study is to explore the clinical significance of Pokemon gene in non-small cell lung cancer(NSCLC). METHODS: Immunohistochemistry was applied to detect the expression of Pokemon protein in 92 cases of NSCLC and 20 cases of paracancerous lung tissues.Correlation between abnormal expression of Pokemon with pathologic characteristics and prognosis of NSCLC was analyzed. RESULTS: Pokemon was not expressed in paracancerous lung tissues and was found in 66 of 92(71.7%) cases of lung cancer tissues.Expression of Pokemon was closely related to TNM stages(P=0.011).Survival rate of patients with negative Pokemon expression was significantly higher than that of those with positive Pokemon expression(P=0.0015).Pokemon expression was demonstrated as independent prognostic factor of NSCLC. CONCLUSIONS: Pokemon is expressed in NSCLC and it may be identified as a new diagnostic marker.High expression of Pokemon may indicate poor prognosis of patients with NSCLC.

[mRNA expression of RASSF1A in esophageal squamous cell carcinoma and clinical significance thereof].

OBJECTIVE: To investigate the mRNA expression of the new tumor suppressor gene, RASSF1A, in esophageal squamous cell carcinoma and its biological value. METHODS: Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the mRNA expression of RASSF1A in the tumor tissues, tissues near tumor, and normal tissues, all obtained during operation, from 66 esophageal squamous cell carcinoma patients. RESULTS: The deletion rates of RASSF1A mRNA expression were 42.4% (28/66), 15.2% (10/66), and 0 in the tumor tissues, tissues near tumor, and normal tissues respectively. The deletion rates of RASSF1A mRNA expression in patients with lymph node metastasis was 61.1%, significantly higher than that of the patients without lymph node metastasis (20.0%, chi(2) = 11.323, P < 0.01); The deletion rates of RASSF1A mRNA expression in the esophageal squamous cell carcinoma at the advanced stages (stages III - IV) was 61.5%, significantly higher than that in the esophageal squamous cell carcinoma at the early stages (stages I - II, 30.0%, chi(2) = 6.417, P < 0.01). The deletion rages of RASSF1A mRNA in the highly, moderately, and lowly differentiation tumors were 38.5% (10/26), 38.5% (10/26), and 57.1% (8/14) respectively, However, there was no significant association between the differentiation degree of tumor and the grade the deletion rages of RASSF1A mRNA (chi(2) = 1.576, P = 0.455). CONCLUSION: The deletion of the mRNA expression of RASSF1A, a tumor suppressor gene, may play an important role in the tumorigenesis and influences the prognosis of esophageal squamous cell carcinoma.