scholarly journals CRABP2 involvement in a mechanism of Golgi stress and tumor dry matter in non-small cell lung cancer cells via ER dependent Hippo pathway

2021 ◽  
Author(s):  
Jian-Feng Meng ◽  
Ming-Jie Luo
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
Hippo Pathway ◽  
Small Cell ◽  
Expression Level ◽  
Control Group ◽  
Nsclc Cell Line ◽  
Small Cell Lung ◽  
Lower Expression ◽  
Er Expression

Objective: The paper aimed to explore the mechanism of cellular retinoic acid binding protein 2 (CRABP2) involvement in Golgi stress and tumor dryness in non-small cell lung cancer (NSCLC) cells through the estrogen receptor (ER) dependent Hippo pathway. Methods: Human NSCLC cell line A549 was purchased from ATCC andcultured in RPMI-1640 with 10% FBS. Attractene reagent was used for plasmid transfection. ER (sh) RNA was designed using RNAi Designer. Seventy-six hours after infection, stable cells were obtained after treated with puromycin for 3 weeks. ER silencing cells (with inhibited ER expression) were compared to the control cells (normal cultured NSCLC cell line A549, CRABP2 normal expression). CRABP2 and ER expression levels were detected by RT-PCR. MTT assay was used to detect cell proliferation, and the cell localization of ER and Golgi was observed by confocal microscopy. The invasion and metastasis of cells were analyzed by Boden chamber invasion and migration assays. Western blotting assays was used for detecting the protein expression of E-cadherin, vimentin, ZO-1 protein and epithelial-mesenchymal transition (EMT) related factors. Results: The lower expression level of mRNA was detected in the ER-silencing group compared to the control group (P<0.05). We also found a higher proliferation level of cells, the number of invading and metastatic cells, the expression of vimentin, p-Lats1T1079, Lats1 and p-YAPS127 mRNA in the control group compared to the ER silencing group (P<0.05). And the expression level of protein kinase RNA-like endoplasmic reticulum kinase (PERK), phosphorylate eukaryotic initiation factor 2 (p-eIF2 alpha), activating transcription factor 4 (ATF4) and C/EBP-homologous protein (CHOP) in the control group was higher than that in the ER silencing group (P<0.05). Adversely, a lower expression level of E-cadherin and ZO-1 protein was found in the control group compared to the ER silencing group (P<0.05). Conclusion: The expression of CRABP2 in NSCLC cells was regulated by ER, and cell proliferation and invasion were regulated by the Hippo pathway. At the same time, it was found that decreased expression of CRABP2 enhanced endoplasmic reticulum/Golgi stress response.

Effects of miR-128 on the Proliferation and Apoptosis of Lung Cancer of Non-Small Cell by Regulating Reversion-Inducing Cysteine-Rich Protein with Kazal Motifs (RECK)

2020 ◽  
Vol 10 (8) ◽  
pp. 1102-1108
Author(s):  
Li-Qun Shan ◽  
Hong-Wang Yan ◽  
Hui Lin ◽  
Hong-Xi Yu ◽  
Bo Zhang ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
Small Cell ◽  
Expression Level ◽  
Control Group ◽  
Nsclc Cell Line ◽  
Related Protein ◽  
Expression Levels ◽  
Proliferation And Apoptosis ◽  
Proliferation Ability

The study was aimed to analyse the effects of miR-128 on the proliferation and apoptosis of lung cancer cells of non-small cell by regulating RECK. The expression of miR-128 in non-small cell lung cancer (NSCLC) and its paracancerours tissues was analyzed by sequencing. miR-128 expression level in NSCLC and its paracancerours tissues was detected by qPCR. We over expressed miR-128 by mimics in A549 NSCLC cell line. Establish blank and negative control group. miR-128 and RECK expression level in each group was detected by qPCR. Use MTT assay to test the proliferation ability of each group. The apoptosis level and the level of ROS in each group were tested by hoechst 33258. The expression level of apoptosis-related protein and p38 NF- B signal pathway-related protein in each group was tested by Western blot. The results of sequencing and qPCR showed that compared with the blank control group, the expression level of miR-128 mRNA was significantly higher in the adjacent tissues than in the adjacent tissues (P < 0 05). The expression levels of miR128 mRNA and RECK mRNA in the overexpression group were significantly increased (P <0 05); the cell proliferation ability in the overexpression group was significantly reduced (P < 0 05), and the level of apoptosis was significantly increased (P < 0 05). Overexpression group Caspase-3, Bcl2/Bax, P-p38, NF-B expression levels were significantly increased (P < 0 05). MiR-128 increases ROS level in NSCLC cells, inhibits cell proliferation and promotes cell apoptosis by regulating RECK and acting on p38 NF-κB signaling pathway.

scholarly journals Nicotine promotes the development of non-small cell lung cancer through activating LINC00460 and PI3K/Akt signaling

2019 ◽  
Vol 39 (6) ◽  
Author(s):  
Hongying Zhao ◽  
Yu Wang ◽  
Xiubao Ren
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
Cell Lines ◽  
Akt Signaling ◽  
Small Cell ◽  
Nsclc Cell ◽  
Expression Level ◽  
In Vitro Experiments ◽  
Small Cell Lung

Abstract Objective: Nicotine, the main ingredient in tobacco, is identified to facilitate tumorigenesis and accelerate metastasis in tumor. Studies in recent years have reported that long intergenic non-protein coding RNA 460 (LINC00460) is strongly associated with lung cancer poor prognosis and nicotine dependence. Nonetheless, it is unclear whether nicotine promotes the development of lung cancer through activation of LINC00460. Methods: We determined that LINC00460 expression in lung cancer tissues and the prognosis in patients with non-small cell lung carcinoma (NSCLC) using Gene Expression Profiling Interactive Analysis (GEPIA) website and The Cancer Genome Atlas (TCGA) database. Through in vitro experiments, we studied the effects of nicotine on LINC00460 in NSCLC cells lines using Cell Counting Kit-8 (CCK-8), transwell test, flow cytometry, quantitative reverse-transcription polymerase chain reaction (qRT-PCR) and Western blot assays. Results: We identified the significant up-regulated expression level of LINC00460 in NSCLC tissues and cell lines, especially, the negative correlation of LINC00460 expression level with overall survival (OS). In in vitro experiments, LINC00460 was overexpressed in NSCLC cell lines under nicotine stimulation. Nicotine could relieve the effect of LINC00460 knockdown on NSCLC cell proliferation, migration and apoptosis. The same influence was observed on PI3K/Akt signaling pathway. Conclusions: In summary, this is the first time to examine the potential roles of LINC00460 in lung cancer cell proliferation, migration and apoptosis induced by nicotine. This may help to develop novel therapeutic strategies for the prevention and treatment of metastatic tumors from cigarette smoke-caused lung cancer by blocking the nicotine-activated LINC00460 pathway.

scholarly journals miRNA-486 and miRNA-499 in human plasma evaluate the clinical stages of lung cancer and play a role as a tumor suppressor in lung tumorigeneisis not pathogenesis

2016 ◽  
Vol 11 (1) ◽  
pp. 264 ◽  
Author(s):  
Youchao Jia ◽  
Aimin Zang ◽  
Yanguang Feng ◽  
Xiao-Fang Li ◽  
Ke Zhang ◽  
...  
Keyword(s):  
Lung Cancer ◽  
Small Cell Lung Cancer ◽  
Cancer Patients ◽  
Cell Lung Cancer ◽  
Statistical Significance ◽  
Small Cell ◽  
Expression Level ◽  
Control Group ◽  
Small Cell Lung ◽  
Lung Cancer Patients

<p class="Abstract">It was aimed to explore the expression level of miRNA-486 and miRNA-499 in the plasma of lung cancer patients and analysis their differences in expre-ssion. The expression level of both miRNA-486 and miRNA-499 in the plasma of non-small cell lung cancer (NSCLC) and small cell lung cancer (SCLC) were lower than that of the control group (p&lt;0.05) and the decrease was more obvious in NSCLC. Compare with the miRNA-499，expression quantity in NSCLC patients plasma. There was statistical significance difference (p&lt;0.05) between III～Ⅳstage and I～II stage. The expression quantity of miRNA in plasma of patients with extensive-stage SCLC was lower than that of patients with limited-stage SCLC (p&lt;0.05). The sensitivity and specificity of plasms miRNA-486 respectively were 88.5% and 83.3%. The expression of miRNA-499 and miRNA-486 in lung cancer patients were up-regulated, and might be closely related to the occurrence and prognosis of lung cancer, and might be used as potential screening and prognosis index for lung cancer.</p><p> </p>

Effect of microRNA-4268 on Proliferation and Apoptosis of Non-Small Cell Lung Cancer Cells Through Regulating Signal Transducer and Activator of Transcription 3 Level

2021 ◽  
Vol 11 (4) ◽  
pp. 725-730
Author(s):  
Lei Han ◽  
Renzhi Yu ◽  
Xin Ni ◽  
Zenglei Zhang
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Mrna Level ◽  
Annexin V ◽  
Small Cell ◽  
Control Group ◽  
Cancer Tissue ◽  
Small Cell Lung

STAT3 is closely related to non-small cell lung cancer (NSCLC). miR-4268 is predicted to regulate STAT3 level by MiRDB analysis. Therefore, our study investigated whether miR-4268 affects NSCLC cells by regulating STAT3. The control group (NC group), miR-4268 Mimics group, and miR-4268 Mimics +pFBD-STAT3 group were set up followed by analysis of miR-4268 and STAT3 mRNA level by QRT-PCR, relationship between miR-4268 and STAT3 by dual fluorescein reporter assay, STAT3 and Tubulin protein level by Western blot, cell proliferation by MTT assay and apoptosis by Annexin V-FITC/PI staining. Compared with normal tissue, miR-4268 expression in cancer tissue was significantly reduced (P <0.01), while STAT3 level was elevated (P <0.01). STAT3 was a target gene of miR-4268. Compared with NC group, STAT3 level was significantly reduced in miR-4268 Mimics group (P <0.01) and increased in miR-4268 Mimics+pFBD-STAT3 group compared with miR-4268 Mimics group (P <0.05). Compared to NC group, miR-4268 Mimics group had reduced cell proliferation and increased cell apoptosis and opposite changes were observed in miR-4268 Mimics+pFBD-STAT3 group which had increased cell proliferation and decreased apoptosis (P < 0.05). miR-4268 regulates STAT3 mRNA level and inhibits NSCLC cell proliferation and promotes apoptosis. However, over-expression of STAT3 can inhibit the effect of miR-4268.

scholarly journals LncRNA SNHG10 is downregulated in non-small cell lung cancer and predicts poor survival

2020 ◽  
Author(s):  
Meng Liang ◽  
Linlin Wang ◽  
Chuanhua Cao ◽  
Shimao Song ◽  
Feng Wu
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Small Cell ◽  
Expression Level ◽  
Small Cell Lung ◽  
Poor Survival ◽  
Tcga Dataset ◽  
Nsclc Patients

Abstract LncRNA SNHG10 has been characterized as an oncogenic lncRNA in liver cancer. By analyzing TCGA dataset we observed the downregulation of SNHG10 in non-small cell lung cancer (NSCLC), indicating its involvement in this disease. We then analyzed the role of SNHG10 in NSCLC.Tumor and paired non-tumor tissues were harvested from 62 NSCLC patients. Expression of SNHG10 and miR-21 in tissues was determined by RT-qPCR. Overexpression of SNHG10 or miR-21 in NSCLC cells was achieved and the interaction between them was analyzed. Cell proliferation was analyzed by CCK-8 assay.In this study we found that SNHG10 is downregulated in cancer tissues of NSCLC patients included in this study. High expression level of SNHG10 predicted favorable survival of NSCLC patients. Levels of miR-21 expression were increased in NSCLC and inversely correlated with SNHG10. In NSCLC cells, SNHG10 overexpression led to increased miR-21 gene methylation and decreased miR-21 expression level. In cell proliferation assay, SNHG10 overexpression attenuated the enhancing effect of miR-21 overexpression on cell proliferation. SNHG10 is downregulated in non-small cell lung cancer and predicts poor survival. It may downregulated miR-21 through methylation to suppress cancer cell proliferation.

Role of IL-6 in neuroendocrine differentiation and chemosensitivity of non-small cell lung cancer

2005 ◽  
Vol 289 (3) ◽  
pp. L438-L445 ◽  
Author(s):  
Kuo-Ting Chang ◽  
Chi-Ying F. Huang ◽  
Chun-Ming Tsai ◽  
Chao-Hua Chiu ◽  
Ying-Yung Lok
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Small Cell ◽  
Cell Counting ◽  
Nsclc Cell Line ◽  
Small Cell Lung ◽  
Transfected Cells ◽  
Mtt Assays

Interleukin-6 (IL-6) has been shown to regulate both growth and neuroendocrine (NE) differentiation in some types of human cancer cells, and erbB2 may be a critical component of IL-6 signaling. Non-small cell lung cancer (NSCLC) tumors that demonstrate NE properties have been suggested to have biological characteristics similar to small cell lung cancers with initial responsiveness to chemotherapy. We investigated whether IL-6 is implicated in the cell growth, NE differentiation, and chemosensitivity of NSCLC-NE cells. NSCLC-NE cells were treated with exogenous IL-6, and a subclone of an IL-6-transfected NSCLC cell line that constitutively expressed IL-6 receptor was also generated. These cells were assessed for cell proliferation by cell counting and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assays, chemosensitivity to cisplatin and etoposide by MTT assays, and NE differentiation by observing morphological changes and immunoblotting for neuron-specific enolase (NSE). The IL-6-treated cells and the IL-6-transfected cells showed enhanced cell proliferation and downregulated NSE expression, but little change in chemosensitivity. In the culture medium, IL-6-transfected cells grew as looser aggregates than the parental cells. IL-6 could not activate the erbB genes. In conclusion, IL-6 can induce cell proliferation and NE dedifferentiation but has little effect on chemosensitivity in IL-6 receptor-expressing NSCLC-NE cells. The status of NSE expression is unlikely to be a crucial factor for chemosensitivity in NSCLC cells.

CALCR knockdown inhibits the development and progression of non-small-cell lung cancer

2021 ◽  
Author(s):  
Tao He ◽  
Feng Ling
Keyword(s):  
Lung Cancer ◽  
Cell Cycle ◽  
Cell Proliferation ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Small Cell ◽  
Expression Level ◽  
Cell Counting ◽  
Small Cell Lung

Abstract G protein-coupled receptors (GPCRs) have been reported to participant in the occurrence and development of a variety of human cancers. CALCR is one of the hundreds of GPCRs, but its expression level and functional importance have never been investigated in non-small-cell lung cancer (NSCLC). In the present study, the protein expression level of CALCR was detected by immunohistochemical staining and western blot analysis. The Celigo cell counting assay was used to assess cell proliferation. Both the wound healing assay and the transwell assay were performed to evaluate cell migration. Flow cytometric analysis was utilized to detect cell apoptosis and cell cycle. A mouse xenograft model was constructed to conduct the in vivo experiments. The results indicated that the CALCR expression was abundantly up-regulated in NSCLC and positively related to tumor infiltrate. Besides, CALCR knockdown could significantly suppress cell proliferation, migration, enhance apoptosis and arrest cell cycle. The in vivo study verified the inhibitory effects of CALCR knockdown on NSCLC tumorigenesis. The abovementioned results provided a reference for the treatment of NSCLC, that was, CALCR knockdown might be a considerable therapeutic strategy.

scholarly journals Midazolam increases cisplatin-sensitivity in non-small cell lung cancer (NSCLC) via the miR-194-5p/HOOK3 axis

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Tingting Sun ◽  
Jing Chen ◽  
Xuechao Sun ◽  
Guonian Wang
Keyword(s):  
Lung Cancer ◽  
Cell Proliferation ◽  
Small Cell Lung Cancer ◽  
Cell Lung Cancer ◽  
Cell Apoptosis ◽  
Cisplatin Resistance ◽  
Small Cell ◽  
Luciferase Reporter ◽  
Small Cell Lung ◽  
Cisplatin Sensitivity

Abstract Backgrounds As previously reported, midazolam anesthesia exerts tumor-suppressing effects in non-small cell lung cancer (NSCLC), but the regulating effects of this drug on cisplatin-resistance in NSCLC have not been studied. Thus, we designed this study to investigate this issue and preliminarily delineate the potential molecular mechanisms. Methods We performed MTT assay and trypan blue staining assay to measure cell proliferation and viability. Cell apoptosis was examined by FCM. qRT-PCR and immunoblotting were performed to determine the expression levels of genes. The targeting sites between genes were predicted by bioinformatics analysis and were validated by dual-luciferase reporter gene system assay. Mice tumor-bearing models were established and the tumorigenesis was evaluated by measuring tumor weight and volume. Immunohistochemistry (IHC) was used to examine the pro-proliferative Ki67 protein expressions in mice tumor tissues. Results The cisplatin-resistant NSCLC (CR-NSCLC) cells were treated with high-dose cisplatin (50 μg/ml) and low-dose midazolam (10 μg/ml), and the results showed that midazolam suppressed cell proliferation and viability, and promoted cell apoptosis in cisplatin-treated CR-NSCLC cells. In addition, midazolam enhanced cisplatin-sensitivity in CR-NSCLC cell via modulating the miR-194-5p/hook microtubule-tethering protein 3 (HOOK3) axis. Specifically, midazolam upregulated miR-194-5p, but downregulated HOOK3 in the CR-NSCLC cells, and further results validated that miR-194-5p bound to the 3’ untranslated region (3’UTR) of HOOK3 mRNA for its inhibition. Also, midazolam downregulated HOOK3 in CR-NSCLC cells by upregulating miR-194-5p. Functional experiments validated that both miR-194-5p downregulation and HOOK3 upregulation abrogated the promoting effects of midazolam on cisplatin-sensitivity in CR-NSCLC cells. Conclusions Taken together, this study found that midazolam anesthesia reduced cisplatin-resistance in CR-NSCLC cells by regulating the miR-194-5p/HOOK3 axis, implying that midazolam could be used as adjuvant drug for NSCLC treatment in clinical practices.

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