scholarly journals GC Content of Early Metazoan Genes and Its Impact on Gene Expression Levels in Mammalian Cell Lines

2018 ◽  
Vol 10 (3) ◽  
pp. 909-917 ◽  
Author(s):  
Ismail Sahin Gul ◽  
Jens Staal ◽  
Paco Hulpiau ◽  
Evi De Keuckelaere ◽  
Kai Kamm ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Gc Content ◽  
Expression Levels ◽  
Mammalian Cell Lines ◽  
Early Metazoan ◽  
Gene Expression Levels

scholarly journals Placing the RPL32 Promoter Upstream of a Second Promoter Results in a Strongly Increased Number of Stably Transfected Mammalian Cell Lines That Display High Protein Expression Levels

2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
F. Hoeksema ◽  
K. Hamer ◽  
M. Siep ◽  
J. A. Verhees ◽  
A. P. Otte
Keyword(s):  
Protein Expression ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Small Region ◽  
High Protein ◽  
Expression Levels ◽  
Mammalian Cell Lines ◽  
Selection Systems ◽  
In Cis ◽  
Gene Expression Levels

The use of high stringency selection systems commonly results in a strongly diminished number of stably transfected mammalian cell lines. Here we placed twelve different promoters upstream of an adjacent primary promoter and tested whether this might result in an increased number of colonies; this is in the context of a stringent selection system. We found that only the promoter of the human ribosomal protein, RPL32, induced a high number of colonies in CHO-DG44 cells. This phenomenon was observed when the RPL32 promoter was combined with the CMV, SV40, EF1-α, and the β-actin promoters. In addition, these colonies displayed high protein expression levels. The RPL32 promoter had to be functionally intact, since the deletion of a small region upstream of the transcription start site demolished its positive action. We conclude that adding the RPL32 promoter to an expression cassette in cis may be a powerful tool to augment gene expression levels.

scholarly journals The Effect of Freeze-Thaw Cycles on Gene Expression Levels in Lymphoblastoid Cell Lines

PLoS ONE ◽  
2014 ◽  
Vol 9 (9) ◽  
pp. e107166 ◽  
Author(s):  
Minal Çalışkan ◽  
Jonathan K. Pritchard ◽  
Carole Ober ◽  
Yoav Gilad
Keyword(s):  
Gene Expression ◽  
Cell Lines ◽  
Lymphoblastoid Cell Lines ◽  
Expression Levels ◽  
Freeze Thaw ◽  
Gene Expression Levels

Development of myeloid-derived suppressor cells (MDSC) targeted therapy for the treatment of cisplatin-resistant bladder cancer.

2019 ◽  
Vol 37 (7_suppl) ◽  
pp. 415-415
Author(s):  
Yuji Takeyama ◽  
Minoru Kato ◽  
Yasuomi Shimizu ◽  
Kosuke Hamada ◽  
Taro Iguchi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bladder Cancer ◽  
Cell Lines ◽  
Cancer Cell ◽  
Suppressor Cells ◽  
Bladder Cancer Cell ◽  
Myeloid Derived Suppressor Cells ◽  
Expression Levels ◽  
Tumor Bearing ◽  
Gene Expression Levels

415 Background: The emergence of immune checkpoint inhibitors (ICI) has brought hope for cure and survival for those suffering from various cancers, including bladder cancer. However, the response rate of ICI monotherapy is modest, and recent reports indicate that myeloid-derived suppressor cells (MDSC) might play a role in the resistant mechanism of ICI. In this study, we assess the effect of chemokine signal on the proliferation of bladder cancer and investigate whether MDSC could be a new target for the treatment of cisplatin-resistant bladder cancer. Methods: We established a cisplatin resistant strain (MB49R) of mice bladder cancer cell line MB49, and examined the alteration of the expression levels of inflammatory chemokines by chemokine array. Next, we isolated MDSCs from spleen and tumor in tumor-bearing mice to examine gene expression levels of chemokine receptors (CXCR2 and CCR2) and immunosuppression genes (Arg-1 and iNOS). Furthermore, we assessed the efficacy of CDDP, α-PD-L1 and chemokine antagonists against the proliferation of tumors in MB49 and MB49R xenograft models. Results: Expression levels of CCL2 and CXCL1/2, which are involved in the migration of MDSC, were significantly increased in the culture supernatant of MB49R compared to those in MB49 cell lines. This result was confirmed by real-time RT-qPCR of tumor extract, and this increase was also observed in human bladder cancer cell lines (T24 and T24R). CXCR2 and CCR2 were highly expressed in PMN-MDSC and M-MDSC, respectively, which were isolated from spleen or tumors in tumor-bearing mice, and gene expression levels of Arg-1 and iNOS were dramatically increased in M-MDSCs from the tumor tissues compared to those from spleen. Also, analysis by flow cytometry revealed that PMN-MDSC dramatically decreased in MB49R compared to parental MB49 tumors, while the proportion of M-MDSC was not changed in MB49R, which indicates that M-MDSC could be a target for the treatment of CDDP resistant bladder cancer. Conclusions: The results in the present study might indicate that the combination treatment with ICI and MDSC-targeting therapy could be an option for the treatment of cisplatin-resistant bladder cancer.

scholarly journals Impact of metallothionein-knockdown on cisplatin resistance in malignant pleural mesothelioma

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Sabrina Borchert ◽  
Pia-Maria Suckrau ◽  
Robert F. H. Walter ◽  
Michael Wessolly ◽  
Elena Mairinger ◽  
...  
Keyword(s):  
Gene Expression ◽  
Protein Expression ◽  
Cell Line ◽  
Cell Lines ◽  
Cisplatin Resistance ◽  
Resistance Mechanism ◽  
Pleural Mesothelioma ◽  
Expression Levels ◽  
Gene Expression Levels

Abstract Malignant pleural mesothelioma (MPM) is a rare, but aggressive tumor with dismal prognosis. Platinum-based chemotherapy is regularly used as part of multimodality therapy. The expression of metallothioneins (MT) has been identified as a reason for cisplatin resistance, which often leads to early therapy failure or relapse. Thus, knockdown of MT expression may improve response to cisplatin treatment. The MT gene- and protein expression of the MPM-cell lines MSTO-211H, NCI-H2052 and NCI-H2452 and the human fibroblast cell line MRC-5, as well as their sensitivity to cisplatin treatment have been evaluated. Knockdown of MT1A, 1B and 2A expression was induced by RNA interference. MT expression was measured using quantitative real-time PCR. An in vitro Assay based on enzyme activity was used to detect cell viability, necrosis and apoptosis before and after incubation with cisplatin. MT2A gene expression could be detected in all MPM cell lines, showing the highest expression in NCI-H2452 and NCI-H2052, whereas gene expression levels of MT1A and MT1B were low or absent. The immunohistochemically protein expression of MT-I/II reflect MT2A gene expression levels. Especially for MSTO-211H cell presenting low initial MT2A levels, a strong induction of MT2A expression could be observed during cisplatin treatment, indicating a cell line-specific and platin-dependent adaption mechanism. Additionally, a MT2A-dependent cellular evasion of apoptosis during cisplatin could be observed, leading to three different MT based phenotypes. MSTO-211H cells showed lower apoptosis rates at an increased expression level of MT2A after cisplatin treatment (from sixfold to fourfold). NCI-H2052 cells showed no changes in MT2A expression, while apoptosis rate is the highest (8–12-fold). NCI-H2452 cells showed neither changes in alteration rate of MT2A expression nor changes in apoptosis rates, indicating an MT2A-independent resistance mechanism. Knockdown of MT2A expression levels resulted in significantly induced apoptotic rates during cisplatin treatment with strongest induction of apoptosis in each of the MPM cell lines, but in different markedness. A therapeutic meaningful effect of MT2A knockdown and subsequent cisplatin treatment could be observed in MSTO-211H cells. The present study showed MT2A to be part of the underlying mechanism of cisplatin resistance in MPM. Especially in MSTO-211H cells we could demonstrate major effects by knockdown of MT2A expression, verifying our hypothesis of an MT driven resistance mechanism. We could prove the inhibition of MT2A as a powerful tool to boost response rates to cisplatin-based therapy in vitro. These data carry the potential to enhance the clinical outcome and management of MPM in the future.

The Gene Expression of Coagulation Factor VIII in Mammalian Cell Lines

1999 ◽  
Vol 95 (2) ◽  
pp. 105-115 ◽  
Author(s):  
Chun Chen ◽  
Xiang-Dong Fang ◽  
Jiang Zhu ◽  
Xiang-Fu Wu ◽  
Zu-Chuan Zhang ◽  
...  
Keyword(s):  
Gene Expression ◽  
Factor Viii ◽  
Cell Lines ◽  
Mammalian Cell ◽  
Coagulation Factor ◽  
Coagulation Factor Viii ◽  
Mammalian Cell Lines

scholarly journals Evaluation of retinoblastoma (Rb) and protein-53 (p53) gene expression levels in breast cancer cell lines (MCF-7) induced with some selected cytotoxic plants

2013 ◽  
Vol 5 (7) ◽  
pp. 120-126 ◽  
Author(s):  
Titilola Aderonke Samuel ◽  
Ayorinde Babatunde James ◽  
Temitope Adesola Oshodi ◽  
Uchennaya Okereke Odii ◽  
Innocent Chidume ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Cell Lines ◽  
Cancer Cell ◽  
Breast Cancer Cell ◽  
P53 Gene ◽  
Breast Cancer Cell Lines ◽  
Expression Levels ◽  
Gene Expression Levels ◽  
Mcf 7
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