OPN-a Splicing Variant Expression in Non-small Cell Lung Cancer and its Effects on the Bone Metastatic Abilities of Lung Cancer Cells In Vitro

Hao, Chengcheng; Cui, Yuxin; Hu, Mu; Zhi, Xiao; Zhang, Lijian; Li, Wenbin; Wu, Wei; Cheng, Shan; Jiang, Wen Guo

doi:10.21873/anticanres.11561

Cited by 15 publications

(17 citation statements)

References 28 publications

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“…Osteopontin (OPN) is a multifunctional, secretory, phosphorylated glycoprotein that can promote cell adhesion and migration. In recent years, OPN has been identified as a critical protein participating in the malignant transformation of tumor cells (12,13). Thus, changes in the expression of MMPs (particularly, MMP-2, MMP-7 and MMP-9), OPN and the NF-κB signaling pathway were evaluated in the current study.…”

Section: Bcsc-1 Suppresses Human Breast Cancer Metastasis By Inhibitimentioning

confidence: 99%

BCSC-1 suppresses human breast cancer metastasis by inhibiting NF-κB signaling

Chen

Guo

et al. 2018

Int J Oncol

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Breast cancer suppressor candidate-1 (BCSC-1; also termed von Willebrand factor A domain containing 5A and LOH11CR2A) is a newly identified candidate tumor suppressor gene that has been implicated in several types of cancer in previous studies. However, there have been few reports about the association between BCSC-1 and human breast cancer in recent years. In the present study, the expression of BCSC-1 in breast cancer was determined by immunohistochemistry (IHC) staining of tissue microarrays and clinical tissue specimens. Subsequently, BCSC-1 gene expression was evaluated in different breast cancer cell lines by quantitative polymerase chain reaction and the MDA-MB-231 cell line was selected for further use in subsequent experiments, due to its low BCSC-1 expression. An MDA-MB-231 cell line with stable overexpression of BCSC-1 was established through transfection with plasmid containing the BCSC-1 gene, and then screening for G418 resistance. Wound-healing, migration and invasion assays were conducted to detect the effect of BCSC-1 on MDA-MB-231 cells. Furthermore, changes in matrix metalloproteinases (MMPs), osteopontin (OPN) and the nuclear factor-κB (NF-κB) pathway were detected in the current study. Additionally, stable silencing of BCSC-1 expression in MCF-7 cells was performed using a lentivirus. The results of IHC indicated that BCSC-1 is expressed at low levels in breast cancer tissues compared with in normal breast tissue. Results of the wound healing, migration and invasion assays demonstrated that BCSC-1 overexpression reduced the metastasis ability of MDA-MB-231 cells in vitro. Further research confirmed that the BCSC-1 overexpression reduced the expression levels of MMP7, MMP9 and OPN, and the phosphorylation of NF-κB p65. Furthermore, inhibition of BCSC-1 via lentivirus-mediated RNA interference revealed that the downregulation of BCSC-1 increased the invasive ability of MCF-7 cells. In summary, the results demonstrated that BCSC-1 is expressed at low levels in breast cancer tissues, and that it can suppress human breast cancer cell migration and invasion, potentially altering the expression of MMP7, MMP9, OPN, and the activity of the NF-κB pathway. Therefore, BCSC-1 may be useful as a biomarker for the treatment of breast cancer in the future.

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Section: Bcsc-1 Suppresses Human Breast Cancer Metastasis By Inhibitimentioning

confidence: 99%

BCSC-1 suppresses human breast cancer metastasis by inhibiting NF-κB signaling

Chen

Guo

et al. 2018

Int J Oncol

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“…Western blot analyses were performed as earlier described [9]. Briefly, samples of cell lysates were…”

Section: Western Blottingmentioning

confidence: 99%

“…OPNa is regarded as the full-length wild type form. OPNb and OPNc are the mutually excluded splicing isoforms, where the former lacks of exon 5 and the latter is without exon 4 [8].The precise regulation and exact functions of different OPN-SIs remained to be obscure, except from recent reports where certain OPN-SI was suggested to be more potent in promoting the development of tumor malignant phenotypes [9,10].Among the known OPN-SIs, OPNc was revealed with most importance of clinical values. OPNc was upregulated in ovarian, breast and pancreatic cancers, which correlated with the histological grade and TNM stage, and well predicted tumor recurrence or metastasis [11][12][13].In CRC patients, OPN protein levels wereincreased in both plasma and tissues, which in negative correlation with the survival [14,15].However, the roles ofOPN-SIs, as well as the change in levels following 5-FU treatments were not fully characterized.…”

Section: Introductionmentioning

confidence: 99%

Epigenetic regulation of osteopontin splicing isoform c defines its role as a microenvironmental factor to promote the survival of colon cancer cells from 5-FU treatment

Chang

Huang

Niu

et al. 2020

Preprint

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Background: Drug resistance to 5-fluorouracil (5-FU) and recurrence after chemotherapy in colorectal cancer remain a challenge to be resolved for the improvement of patient outcomes. It is recognized that a variety of secretory proteins released from the tumor cells exposed to chemo-drugsinto the tumor microenvironment (TME) contributedto the cell-to-cell communication, and altered the drug sensitivity.One of these important factors is osteopontin (OPN), which exists in several functional forms from alternative splicing and post-translational processing. In colon cancer cells, increased total OPN expression was observed during the progression of tumors, however, the exact role and regulation of the OPN splicing isoforms was not well understood. Methods: We assayed precisely the abundance of majorOPN splicing isoformsunder5-FU treatmentsincolon cancer cell lineswith different sensitivities to 5-FU, and also evaluated the effects of the condition medium from OPNsplicing isoforms overexpressed cells on cell functions. The methods of nuclear calcium reporter assays and ChIP (chromatin immunoprecipitation) assays were used to investigate the molecular mechanism underlining the production of OPN isoforms.Results: We discovered that OPNc was a most increased splicing isoform to a significant abundance following 5-FU treatment of colon cancer cells. OPNc as a secretory protein in the conditioned medium exerted a more potent effect to promote cell survival in 5-FU than other OPN isoforms. The kinetic response of nuclear calcium signalscouldbe used to indicate an immediate effect of the conditioned medium containing OPNc and other isoforms. MeCP2 was identified to regulate the splicing of opn gene, where the phosphorylation of MeCP2 at S421 site, possibly by CAMKII, was required. Conclusions: The results demonstrated that the production of OPNc was highly controlled under epigenetic regulations, where MeCP2 and the activation of nuclear calcium signaling were involved. It was also suggested that OPNc could transmit the stress signal of cells upon chemotherapy in TME and promoted the survival of adjacent colon cancer cells.

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“…Western blot analyses were performed as earlier described [9]. Brie y, samples of cell lysates were prepared and separated by 10% SDS-polyacrylamide gel electrophoresis (SDS-PAGE), then transferred onto polyvinylidene uoride (PVDF) lters.…”

Section: Western Blottingmentioning

confidence: 99%

Epigenetic regulation of osteopontin splicing isoform c defines its role as a microenvironmental factor to promote the survival of colon cancer cells from 5-FU treatment

Chang

Huang

Niu

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

Abstract Background: Drug resistance to 5-fluorouracil (5-FU) and recurrence after chemotherapy in colorectal cancer remain a challenge to be resolved for the improvement of patient outcomes. It is recognized that a variety of secretory proteins released from the tumor cells exposed to chemo-drugs into the tumor microenvironment (TME) contributed to the cell-to-cell communication, and altered the drug sensitivity. One of these important factors is osteopontin (OPN), which exists in several functional forms from alternative splicing and post-translational processing. In colon cancer cells, increased total OPN expression was observed during the progression of tumors, however, the exact role and regulation of the OPN splicing isoforms was not well understood.Methods: We assayed precisely the abundance of major OPN splicing isoforms under 5-FU treatments in colon cancer cell lines with different sensitivities to 5-FU, and also evaluated the effects of the condition medium from OPN splicing isoforms overexpressed cells on cell functions. The methods of nuclear calcium reporter assays and ChIP (chromatin immunoprecipitation) assays were used to investigate the molecular mechanism underlining the production of OPN isoforms.Results: We discovered that OPNc was a most increased splicing isoform to a significant abundance following 5-FU treatment of colon cancer cells. OPNc as a secretory protein in the conditioned medium exerted a more potent effect to promote cell survival in 5-FU than other OPN isoforms. The kinetic response of nuclear calcium signals could be used to indicate an immediate effect of the conditioned medium containing OPNc and other isoforms. Methyl-CpG binding protein 2 (MeCP2) was identified to regulate the splicing of opn gene, where the phosphorylation of MeCP2 at S421 site, possibly by calmodulin dependent protein kinase II (CaMKII) was required.Conclusions: The results demonstrated that the production of OPNc was highly controlled under epigenetic regulations, where MeCP2 and the activation of nuclear calcium signaling were involved. It was also suggested that OPNc could transmit the stress signal of cells upon chemotherapy in TME and promoted the survival of adjacent colon cancer cells.

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OPN-a Splicing Variant Expression in Non-small Cell Lung Cancer and its Effects on the Bone Metastatic Abilities of Lung Cancer Cells In Vitro

Abstract: OPN-a may represent a bone metastatic factor in human lung cancer, as well as a potential therapy target.

Cited by 15 publications

References 28 publications

BCSC-1 suppresses human breast cancer metastasis by inhibiting NF-κB signaling

BCSC-1 suppresses human breast cancer metastasis by inhibiting NF-κB signaling

Epigenetic regulation of osteopontin splicing isoform c defines its role as a microenvironmental factor to promote the survival of colon cancer cells from 5-FU treatment

Epigenetic regulation of osteopontin splicing isoform c defines its role as a microenvironmental factor to promote the survival of colon cancer cells from 5-FU treatment

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