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

scholarly journals P5CR1 protein expression and the effect of gene-silencing on lung adenocarcinoma

PeerJ ◽  
2019 ◽  
Vol 7 ◽  
pp. e6934 ◽  
Author(s):  
Yang She ◽  
Aiyou Mao ◽  
Feng Li ◽  
Xiaobin Wei
Keyword(s):  
Protein Expression ◽  
Lung Adenocarcinoma ◽  
Gene Silencing ◽  
Therapeutic Target ◽  
Biological Behavior ◽  
Pathological Grade ◽  
Adenocarcinoma Cells ◽  
Cisplatin Sensitivity ◽  
Lung Adenocarcinoma Cells ◽  
And Gender

The present study aimed to investigate the expression of pyrroline-5-carboxylate reductase 1 (P5CR1) protein in lung adenocarcinoma and paracancerous tissues and to explore the effect of silencing the encoding gene PYCR1 on the proliferation, migration, invasion, and cisplatin sensitivity in lung adenocarcinoma cells, thereby providing a novel therapeutic target for the treatment of the disease. Immunohistochemistry staining was used to detect the P5CR1 protein expression in lung adenocarcinoma and paracancerous tissues, and statistical analysis evaluated the correlation between P5CR1 protein expression and gender, age, tissue part, or pathological grade. The CCK8 assay was performed to detect the proliferation and cisplatin sensitivity, while the effect of PYCR1 on the migration and invasion of lung adenocarcinoma cells was detected by scratch test and transwell chamber assay. The findings demonstrated that the P5CR1 protein expression was significantly elevated in lung adenocarcinoma tissues and correlated with the pathological grade, whereas no significant correlation was established between the protein expression and gender, age, or tissue part. Furthermore, after PYCR1 gene silencing, the proliferation and invasion were significantly suppressed, while the sensitivity to cisplatin was significantly enhanced. Therefore, it can be speculated that the PYCR1 gene affects the biological behavior of lung adenocarcinoma and cisplatin resistance, serving as a potential therapeutic target for lung adenocarcinoma.

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.

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 PFKFB4 promotes lung adenocarcinoma progression via phosphorylating and activating transcriptional coactivator SRC-2

2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Jiguang Meng ◽  
Xuxin Chen ◽  
Zhihai Han
Keyword(s):  
Lung Adenocarcinoma ◽  
Transcriptional Activity ◽  
Cell Counting ◽  
Migration And Invasion ◽  
Transcriptional Coactivator ◽  
Downstream Target ◽  
Steroid Receptor Coactivator ◽  
Adenocarcinoma Cells ◽  
Family Protein ◽  
Lung Adenocarcinoma Cells

Abstract Background To investigate the role and its potential mechanism of 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase 4 (PFKFB4) in lung adenocarcinoma. Methods Co-immunoprecipitation was performed to analyze the interaction between PFKFB4 and SRC-2. Western blot was used to investigate the phosphorylation of steroid receptor coactivator-2 (SRC-2) on the condition that PFKFB4 was knockdown. Transcriptome sequencing was performed to find the downstream target of SRC-2. Cell Counting Kit-8 (CCK-8) assay, transwell assay and transwell-matrigel assay were used to examine the proliferation, migration and invasion abilities in A549 and NCI-H1975 cells with different treatment. Results In our study we found that PFKFB4 was overexpressed in lung adenocarcinoma associated with SRC family protein and had an interaction with SRC-2. PFKFB4 could phosphorylate SRC-2 at Ser487, which altered SRC-2 transcriptional activity. Functionally, PFKFB4 promoted lung adenocarcinoma cells proliferation, migration and invasion by phosphorylating SRC-2. Furthermore, we identified that CARM1 was transcriptionally regulated by SRC-2 and involved in PFKFB4-SRC-2 axis on lung adenocarcinoma progression. Conclusions Our research reveal that PFKFB4 promotes lung adenocarcinoma cells proliferation, migration and invasion via enhancing phosphorylated SRC-2-mediated CARM1 expression.

scholarly journals PD-L1 lncRNA splice isoform promotes lung adenocarcinoma progression via enhancing c-Myc activity

2021 ◽  
Vol 22 (1) ◽  
Author(s):  
Shuang Qu ◽  
Zichen Jiao ◽  
Geng Lu ◽  
Bing Yao ◽  
Ting Wang ◽  
...  
Keyword(s):  
Alternative Splicing ◽  
Lung Adenocarcinoma ◽  
Solid Tumors ◽  
Immune Checkpoint ◽  
Checkpoint Inhibition ◽  
Immune Checkpoint Inhibition ◽  
Non Coding Rna ◽  
Adenocarcinoma Cells ◽  
Lung Adenocarcinoma Cells ◽  
Long Non Coding Rna

Abstract Background Although using a blockade of programmed death-ligand 1 (PD-L1) to enhance T cell immune responses shows great promise in tumor immunotherapy, the immune-checkpoint inhibition strategy is limited for patients with solid tumors. The mechanism and efficacy of such immune-checkpoint inhibition strategies in solid tumors remains unclear. Results Employing qRT-PCR, Sanger sequencing, and RNA BaseScope analysis, we show that human lung adenocarcinoma (LUAD) all produce a long non-coding RNA isoform of PD-L1 (PD-L1-lnc) by alternative splicing, regardless if the tumor is positive or negative for the protein PD-L1. Similar to PD-L1 mRNA, PD-L1-lnc in various lung adenocarcinoma cells is significantly upregulated by IFNγ. Both in vitro and in vivo studies demonstrate that PD-L1-lnc increases proliferation and invasion but decreases apoptosis of lung adenocarcinoma cells. Mechanistically, PD-L1-lnc promotes lung adenocarcinoma progression through directly binding to c-Myc and enhancing c-Myc transcriptional activity. Conclusions In summary, the PD-L1 gene can generate a long non-coding RNA through alternative splicing to promote lung adenocarcinoma progression by enhancing c-Myc activity. Our results argue in favor of investigating PD-L1-lnc depletion in combination with PD-L1 blockade in lung cancer therapy.

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