scholarly journals Study on the Mechanism of Cancer/Testicular Antigen FSIP1 in Regulating Autophagy and Promoting Survival of Lung Cancer Cells

2021 ◽  
Vol 2 (1) ◽  
Author(s):  
Xiaoping Zhu ◽  
Yingping Song ◽  
Wei Wang ◽  
Weiqi Yang
Keyword(s):  
Gene Expression ◽  
Quantitative Analysis ◽  
Lung Adenocarcinoma ◽  
Actinomycin D ◽  
Normal Lung ◽  
Time Points ◽  
Protein Levels ◽  
Adenocarcinoma Cells ◽  
Significant Difference ◽  
Lung Adenocarcinoma Cells

Objective — To investigate the molecular mechanism of FSIP1 gene expression and autophagy inhibition in patients with lung adenocarcinoma. Methods — TTP expression in lung adenocarcinoma H1299, H1975 and normal lung forming cells was detected by QRT-PCR and Western Bloting. Quantitative analysis and Western Bloting were performed by transfecting pCMV-FSIPI at 24h, 48h and 72h respectively to analyze the expression of autophagy related factors like P62, LC3II/LCI and BecIinl. The transient transfection of overexpressed and empty plasmid was performed by adding 10 mg/ml of actinomycin D. After the termination of this transcription, RNA extraction was performed at different time points to detect the expression of Beclin1m RNA at different time points. After adding TNF-a by transfection plasmid, lung adenocarcinoma cells were divided into different groups, including no-load group, FSIPI group, no-load + TNF-A group, and FSIPI + TNF-a group. The gene expression of P50, c-Rel and NF-KBp65 in the nucleus was analyzed by immunofluorescence and Western Bloting. Lung adenocarcinoma was divided into Ikba-mut no-load group, FSIPI no-load group, 1Ikba-mut group, FSIPI group and FSIPI+ Ikba-mut group. FSIPI expression and autophagy gene expression were detected by quantitative analysis and Western Bloting. Results — In lung adenocarcinoma cells, FSIP1 RNA and protein levels were RELATIVELY low, and autophagy Becline and LC 3II/I had lower RNA and protein levels after overexpression of FSIPI compared with the no-load group. Transcription is terminated by the addition of actinomycin D. There was no significant difference in the expression of the autophagy-related gene BEC LINEM between FSIP1 and the no-load group. After overexpression of FSIPI, according to Western Bloting results, nuclear C-Rel and P65 proteins were less than those in the no-load +TNF-a group and the no-load group, whileP50 protein did not change significantly. Combined with immunofluorescence studies, it was found that the expressions of c-Rel and P65 were significantly decreased after FSIP1 overexpression compared with the no-load group, while the expression of P50 was not significantly changed. Conclusion — FSIP1 is usually low expressed in lung adenocarcinoma. Overexpression of FSIP1 can effectively inhibit the nuclear metastasis of C-Rel and NF-KBp65, thus inhibiting autophagy of the cells.

scholarly journals RNAi Targeting-STIL Suppresses Cell Growth and Induces Apoptosis of Lung Cancer NCI-H1299 Cells via Inactivation of Akt/SAPK/TAK1 Pathways

2021 ◽  
Author(s):  
Hailong Li ◽  
Rong Niu ◽  
Yi Zhang ◽  
Yanmei Song ◽  
Jing Wang ◽  
...  
Keyword(s):  
Cell Growth ◽  
Lung Adenocarcinoma ◽  
Colony Formation ◽  
Normal Lung ◽  
Antibody Array ◽  
Signal Pathways ◽  
Pathological Characteristic ◽  
Adenocarcinoma Cells ◽  
Inhibit Cell Growth ◽  
Lung Adenocarcinoma Cells

Abstract Background: Gradually emerged studies demonstrated that SCL/TAL1 interrupting locus (STIL or SIL) is upregulated in multiple kinds of fatal tumors; at present, there is no clean understanding about the role of STIL in lung adenocarcinoma cells. This study aimed to discover the significance of STIL in lung adenocarcinoma, so as to find a potential gene target for diagnosis and therapy. Methods: STIL expression in lung adenocarcinoma tissue and clinical pathological characteristic was analyzed using the online databases, UALCAN and GEPIA. Lentivirus STIL-shRNA was manufactured and transducted into lung adenocarcinoma cells to seek and analyze the effects on tumor phenotype. The cell proliferation was assessed using Cellomics Array Scan imaging assay, and colony-formation assay, respectively. The apoptosis was detected by flow cytometry assay. Moreover, the antibody array of PathScan Cancer Phenotype, PathScan stress and apoptosis pathway was used to explore relevant molecular mechanisms following STIL knockdown in NCI-H1299 cells. Results: The clinical pathological characteristic assay showed that STIL is upregulated in lung adenocarcinoma tissues, and this trend was associated with cancer stage1 and histological subtypes. Silencing experiment showed that downregulation of STIL could inhibit cell growth and colony formation, induce cell apoptosis, and a G2 phase arrest effect significantly, and antibody array detection revealed that p-Bad were upregulated, and p-Akt, p-Bad, p-HSP27, p-SAPK/JNK, p-TAK1, Vimentin, CD45, PCNA and Ki-67 were downregulated significantly after STIL silenced in NCI-H1299 cells.Conclusions: In conclusion, STIL is overexpressed in lung adenocarcinoma tissues compared with normal lung tissue. Knockdown of STIL could inhibit cell growth and colony formation ability, promote apoptosis via Akt/SAPK/TAK1 signal pathways inactivation.

scholarly journals Radiosensitization of non-small cell lung carcinoma by EGFR inhibition

2014 ◽  
Vol 29 (3) ◽  
pp. 233-241 ◽  
Author(s):  
Otilija Keta ◽  
Tanja Bulat ◽  
Lela Koricanac ◽  
Jelena Zakula ◽  
Giacomo Cuttone ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Small Cell Lung Carcinoma ◽  
Combined Treatments ◽  
Cell Lung Carcinoma ◽  
Cell Inactivation ◽  
Egfr Inhibition ◽  
Time Points ◽  
Adenocarcinoma Cells ◽  
Lung Adenocarcinoma Cells

Molecular targeted cancer therapy is a promising treatment strategy. Considering the central role of the epidermal growth factor receptor in cell proliferation and survival, there are indications that targeted agents like tyrosine kinase inhibitors, i. e., erlotinib, may enhance the antitumor treatment by radiation. The aim of this study is to analyze the inactivation effects of g-rays and to test the radiosensitizing potential of erlotinib on human lung adenocarcinoma cells in vitro. Irradiations were performed with doses ranging from 1 Gy to 8 Gy. In order to increase the radiosensitivity of CRL-5876 lung adenocarcinoma cells, the cells were treated with a clinically relevant concentration of 2 ?M erlotinib. The effects of single and combined treatments were monitored using clonogenic survival, cell viability and proliferation assays at different time points. For the detection and visualization of the phosphorylated histone H2AX (?-H2AX), an important biological marker of DNA double-strand break formation, fluorescence immunocytochemistry, was performed. The response to the treatment was monitored at four time points: 30 min, 2, 6, and 24 h. Irradiations with g-rays resulted in significant cell inactivation regarding all analyzed biological endpoints. Combined treatments revealed consistent cell inactivation. Moreover, compared to g-rays alone, elevated levels of g-H2AX foci were observed after pretreatment with erlotinib, indicating radiosensitization through impaired DNA repair.

scholarly journals ID2 Inhibits Lung Adenocarcinoma Cell Malignant Behaviors Through Inhibiting the Activation of the PI3K/AKT/mTOR Signaling Pathway

2021 ◽  
Author(s):  
Wenzhong Peng ◽  
Jia Chen ◽  
Ruoxi He ◽  
Yongjun Tang ◽  
Juan Jiang ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Candidate Gene ◽  
Signaling Pathway ◽  
Mtor Signaling ◽  
Mtor Signaling Pathway ◽  
Tissue Samples ◽  
Adenocarcinoma Cell ◽  
Protein Levels ◽  
Adenocarcinoma Cells ◽  
Lung Adenocarcinoma Cells

Abstract Background: Lung cancer is the most common cancer and one of the main causes of cancer-related deaths, and it manifests as metastatic disease in most cases. Considering frequent gene mutation and/or signaling deregulation in lung adenocarcinoma, identifying novel factors or agents targeting these signaling pathways might be promising strategies for lung adenocarcinoma therapy. Methods: GEO datasets were analyzed to identify differentially expressed genes (DEGs) in lung adenocarcinoma. The specific effects of candidate gene overexpression or knockdown on lung adenocarcinoma cell phenotypes were examined. Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) are used to connect the genomic and functional information of DEGs. The dynamic effects of candidate gene and signaling pathway agonist on lung adenocarcinoma malignant behaviors were investigated. Finally, clinical lung adenocarcinoma and adjacent non-cancerous tissues were collected and the levels of candidate gene were examined in tissue samples.Results: Inhibitor of DNA binding 2 (ID2) was identified as an aberrantly downregulated gene in lung adenocarcinoma. ID2 overexpression suppressed lung adenocarcinoma cell viability, colony formation capacity, and migration. ID2 overexpression also reduced the protein levels of N-cadherin, MMP2, MMP9, and the phosphorylation of AKT and mTOR. The PI3K/AKT/mTOR signaling agonist exerted opposite effects on lung adenocarcinoma cells to those of ID2 overexpression, and partially reversed the effects of ID2 overexpression. In tissue samples, ID2 protein levels and mRNA expression were also downregulated compared with those in adjacent non-cancerous tissues. Conclusion: ID2 exerts its tumor-suppressive effects on lung adenocarcinoma cell malignant behaviors through inhibiting the activation of the PI3K/AKT/mTOR signaling pathway. Restoring ID2 expression in lung adenocarcinoma cells might improve the curative effect of lung adenocarcinoma therapies.

scholarly journals Gender difference in the activity but not expression of estrogen receptors α and β in human lung adenocarcinoma cells

2006 ◽  
Vol 13 (1) ◽  
pp. 113-134 ◽  
Author(s):  
Susan M Dougherty ◽  
Williard Mazhawidza ◽  
Aimee R Bohn ◽  
Krista A Robinson ◽  
Kathleen A Mattingly ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
Cell Lines ◽  
Human Lung ◽  
Lung Fibroblasts ◽  
Normal Lung ◽  
Adenocarcinoma Cell ◽  
Human Lung Adenocarcinoma ◽  
Estrogen Response ◽  
Adenocarcinoma Cells ◽  
Lung Adenocarcinoma Cells

The higher frequency of lung adenocarcinoma in women smokers than in men smokers suggests a role for gender-dependent factors in the etiology of lung cancer. We evaluated estrogen receptor (ER) α and β expression and activity in human lung adenocarcinoma cell lines and normal lung fibroblasts. Full-length ERα and ERβ proteins were expressed in all cell lines with higher ERβ than ERα. Although estradiol (E2) binding was similar, E2 stimulated proliferation only in cells from females, and this response was inhibited by anti-estrogens 4-hydroxytamoxifen (4-OHT) and ICI 182,780. In contrast, E2 did not stimulate replication of lung adenocarcinoma cells from males and 4-OHT or ICI did not block cell proliferation. Similarly, transcription of an estrogen response element-driven reporter gene was stimulated by E2 in lung adenocarcinoma cells from females, but not males. Progesterone receptor (PR) expression was increased by E2 in two out of five adenocarcinoma cell lines from females, but none from males. E2 decreased E-cadherin protein expression in some of the cell lines from females, as it did in MCF-7 breast cancer cells, but not in the cell lines from males. Thus, ERα and ERβ expression does not correlate with the effect of ER ligands on cellular activities in lung adenocarcinoma cells. On the other hand, coactivator DRIP205 expression was higher in lung adenocarcinoma cells from females versus males and higher in adenocarcinoma cells than in normal human bronchial epithelial cells. DRIP205 and other ER coregulators may contribute to differences in estrogen responsiveness between lung adenocarcinoma cells in females and males.

scholarly journals Effect of shRNA-Mediated Knockdown EBF1 Gene Expression on the Proliferation of Lung Cancer Cell Line A549 In Vitro and In Vivo

2021 ◽  
Author(s):  
Lin WANG ◽  
Ding Li ◽  
Li REN ◽  
Honglei FENG
Keyword(s):  
Gene Expression ◽  
Lung Cancer ◽  
Cell Cycle ◽  
Lung Adenocarcinoma ◽  
A549 Cells ◽  
Adenocarcinoma Cells ◽  
Lung Adenocarcinoma Cells ◽  
Cancer Tissues ◽  
Experimental Group

Abstract BackgroundThe incidence of lung cancer is increasing year by year. The study on the proliferation and metastasis of lung adenocarcinoma cells is of positive significance to improve the prognosis of patients with lung adenocarcinoma, but there is still a lack of more effective treatment for the proliferation and metastasis of lung adenocarcinoma cells .Here we find that a lymphocyte lineage specific transcription factor,EBF1, is frequently expressed in human lung cancer tissues and affects the proliferation of tumor cells , Objective to explore the possible mechanism of affecting the proliferation of lung adenocarcinoma cells.MethodsImmunohistochemistry and PCR were used to detect the expression of EBF1 in lung cancer tissues and lung cancer cell lines. According to the interference RNA (shRNA) sequence designed by our laboratory for EBF1 as the target sequence and a random sequence as the negative control, the recombinant retrovirus was constructed and transfected into A549 cells, which were used as A549-shRNA-EBF1 and A549-shRNA-control of experimental group and control group respectively; Knockdown of EBF1 gene was detected by PCR and Western blot. MTT and BrdU staining were used to detect the effect of EBF1-shRNA on the proliferation of A549 cells in vitro;flow cytometry was used to analyze the cell cycle of each group; subcutaneous inoculation of cells in axilla of nude mice was used to observe the effect of EBF1-shRNA on the tumorigenicity of A549 cells in nude mice; Western blot was used to detect the expression of CDK6, P21 and P27 proteins.ResultsEBF1 was not expressed in stromal cells of adjacent tissues and lung cancer tissues, but in nuclei of NSCLC and SCLC cancer cells. EBF1-shRNA knockdown EBF1 gene expression effectively; knockdown EBF1 gene expression can inhibit the proliferation of A549 cells in vitro and in vivo, and block the cell cycle of experimental group at G1 phase; after knockdown EBF1 gene expression, CDK6 protein expression in experimental group cells decreases, while P21 and P27 protein increase.ConclusionsEBF1-shRNA can inhibit the proliferation of lung adenocarcinoma A549 in vitro and in vivo by blocking cell cycle in G1 phase, which involves the decrease of CDK6 expression and the up-regulation of P21 and P27 expression. This study will supplement the theory that heterotopic expression of hematogenous transcription factors in lung cancer affects tumor proliferation and discover new molecular targets for cancer therapy.

The effect of RNAi-mediated gene silencing on her-2/neu gene expression in lung adenocarcinoma cells

2005 ◽  
Vol 17 (2) ◽  
pp. 84-89 ◽  
Author(s):  
Shu-hua Ren ◽  
Jing-wei Wang ◽  
Ping Qu ◽  
Yi Liu ◽  
Wei Zhang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Lung Adenocarcinoma ◽  
Gene Silencing ◽  
Adenocarcinoma Cells ◽  
Her 2 ◽  
Lung Adenocarcinoma Cells
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