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

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.

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

Planta Medica ◽

10.1055/s-0033-1352052 ◽

2013 ◽

Vol 79 (13) ◽

Cited By ~ 1

Author(s):

TA Samuel ◽

B James ◽

T Oshodi ◽

U Odii ◽

OA Odukoya ◽

...

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

Scientific Reports ◽

10.1038/s41598-020-75807-x ◽

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.

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

Journal of Pharmacognosy and Phytotherapy ◽

10.5897/jpp2013.0270 ◽

2013 ◽

Vol 5 (7) ◽

pp. 120-126 ◽

Cited By ~ 1

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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Clinical drug response can be predicted using baseline gene expression levels and in vitro drug sensitivity in cell lines

Genome Biology ◽

10.1186/gb-2014-15-3-r47 ◽

2014 ◽

Vol 15 (3) ◽

pp. R47 ◽

Cited By ~ 180

Author(s):

Paul Geeleher ◽

Nancy J Cox ◽

R Huang

Keyword(s):

Gene Expression ◽

Cell Lines ◽

Drug Response ◽

Drug Sensitivity ◽

Expression Levels ◽

Clinical Drug ◽

Gene Expression Levels

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Trichostatin A preferentially reverses the upregulation of gene-expression levels induced by gain of chromosome 7 in colorectal cancer cell lines

Genes Chromosomes and Cancer ◽

10.1002/gcc.22505 ◽

2017 ◽

Vol 57 (1) ◽

pp. 35-41

Author(s):

Floryne O. Buishand ◽

Eric Cardin ◽

Yue Hu ◽

Thomas Ried

Keyword(s):

Gene Expression ◽

Colorectal Cancer ◽

Cell Lines ◽

Cancer Cell ◽

Trichostatin A ◽

Chromosome 7 ◽

Colorectal Cancer Cell ◽

Expression Levels ◽

Colorectal Cancer Cell Lines ◽

Gene Expression Levels

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Transcriptional expression of survivin and its splice variants in cervical carcinomas

International Journal of Gynecological Cancer ◽

10.1111/j.1525-1438.2007.00833.x ◽

2007 ◽

Vol 17 (5) ◽

pp. 1092-1098 ◽

Cited By ~ 14

Author(s):

H. Futakuchi ◽

M. Ueda ◽

K. Kanda ◽

K. Fujino ◽

H. Yamaguchi ◽

...

Keyword(s):

Gene Expression ◽

Cell Lines ◽

Splice Variants ◽

Transcriptional Expression ◽

Tissue Samples ◽

Expression Levels ◽

Cervical Carcinomas ◽

Significant Difference ◽

Normal Controls ◽

Gene Expression Levels

The objective of this study was to evaluate transcriptional expression of survivin and the two splice variants (survivin-2B and survivin-ΔEx3) in cervical carcinomas. The gene expression levels of survivin and its splice variants in 11 human cervical carcinoma cell lines and 20 malignant and 12 normal cervical tissue samples were analyzed using quantitative reverse transcription–polymerase chain reaction analysis. Gene expression levels of survivin and survivin-ΔEx3 in cell lines were higher than those in normal cervical tissues (P= 0.0193 and 0.0489). Transcript levels of survivin and survivin-ΔEx3 in carcinoma tissues were also higher than those in normal controls (P= 0.0016 and 0.0011). Gene expression levels of survivin and survivin-ΔEx3 in adenocarcinomas were statistically higher than those in squamous cell carcinomas (P= 0.0260 and 0.0487). There was no significant difference in survivin-2B gene expression between malignant and normal cervical samples or different histologic types. The ratios of survivin-2B/survivin and survivin-ΔEx3/survivin in carcinoma tissues were higher than those in normal controls (P= 0.0288 and 0.0081). Interestingly, the ratio of survivin-2B/survivin was increased in the patients with higher stages and with pelvic lymph node metastasis (P= 0.0205 and 0.0437), respectively. We conclude that survivin and its splice variants might be involved in the pathogenesis and development of cervical carcinomas.

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