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Pankreas Kanseri Hücrelerinde Tripartite Motif-Containing Protein 3 (TRIM3) Gen Ekspresyonunun Araştırılması

Year 2022, , 505 - 509, 27.12.2022
https://doi.org/10.35440/hutfd.1121746

Abstract

Amaç: Pankreas kanseri, tüm kanserler içinde en kötü prognoza sahip olanlar arasındadır. Tripartite Motif-Containing Protein 3 (TRIM3) geni tümör baskılayıcı bir gen olarak kanser hücrelerinin proliferasyonu, migrasyonu ve invazyonunu kontrol ederek tümör baskılayıcı olarak rol oynamaktadır. Bu çalışmanın amacı, AsPC1, BxPC-3 ve PANC-1 pankreas kanseri hücre hatlarındaki TRIM3 geninin mRNA seviyesindeki ekspresyonunu araştırmaktır.

Materyal ve metod: AsPC1, BxPC-3 ve PANC-1 hücre hatları 37˚C’de %5 CO2 içeren ortamda kültüre edildi ve total RNA izolasyonu yapıldı. TRIM3 geni mRNA ekspresyon seviyesi Kantitatif Ters Transkripsiyon PCR (RT-qPCR) metodu ile incelendi. Relatif gen ekspresyon verilerinin analizi 2-∆∆CT metodu kullanılarak yapıldı.

Bulgular: Üç hücre hattında da TRIM3'ün mRNA ekspresyon seviyelerinin çok düşük olduğu tespit edildi. İlaveten kat değişimi hesaplandığında hücre hatları arasında istatistiksel fark gözlenmedi.

Sonuç: TRIM3 geni karsinogenez sürecinde tümör baskılayıcı gen olarak rol oynamaktadır ve kanser hücrelerinde TRIM3 ekspresyonunun azaldığı gösterilmiştir. Literatürdeki diğer kanser türleri ile uyumlu şekilde pankreas kanseri hücrelerinde TRIM3 mRNA ekspresyonunun çok düşük olduğu tespit edilmiştir. Bu çalışma AsPC1, BxPC-3 ve PANC-1 pankreas kanseri hücre hatları ve TRIM3 arasındaki ilişkiyi araştıran tek çalışma olması sebebiyle bundan sonra yapılacak fonksiyonel çalışmalara ışık tutacaktır.

References

  • Siegel RL, Miller KD, Jemal A. Cancer Statistics, 2020. Ca Cancer J Clin. 2020; 70:7-30.
  • Sharma C, Eltawil KM, Renfrew PD, Walsh MJ, Molinari M. Advances in diagnosis, treatment and palliation of pancreatic carcinoma: 1990-2010. World J Gastroenterol 2011; 17(7):867-897.
  • Omura N, Li CP, Li A, Hong SM, Walter K, Jimeno A, et al. Genome-wide profiling of methylated promoters in pancreatic adenocarcinoma. Cancer Biol Ther. 2008; 7(7):1146-1156.
  • Jones S, Hruban RH, Kamiyama M, Borges M, Zhang X, Parsons DW, et al. Exomic Sequencing Identifies PALB2 as a Pancreatic Cancer Susceptibility Gene. Science. 2009; 324(5924): 217-219.
  • Klein AP, Wolpin BM, Risch HA, Stolzenberg-Solomon RZ, Mocci E, Zhang M, et al. Genome-wide meta-analysis identifies five new susceptibility loci for pancreatic cancer. Nature Communications. 2018; 9:556-566.
  • Wolpin BM, Rizzato C, Kraft P, Kooperberg C, Petersen GM, Wang Z, et al. Genome-wide association study identifies multiple susceptibility loci for pancreatic cancer. Nature Genetics. 2014; doi:10.1038/ng.3052.
  • Meroni G, Diez-Roux G. TRIM/RBCC, a novel class of ‘single protein RING finger’ E3 ubiquitin ligases. Bioessays. 2005; 27(11):1147-1157.
  • Huang XQ, Zhang XF, Xia JH, Chao J, Pan QZ, Zhao JJ, et al. Tripartite motif containing 3 (TRIM3) inhibits tumor growth and metastasis of liver cancer. Chin J Cancer 2017; 36: 77.
  • Wang M, Wu J, Guo Y, Chang X, Cheng T, The tripartite motif-containing protein 3 on the proliferation and cytokine secretion of rheumatoid arthritis fibroblast-like synoviocytes. Mol. Med. Rep. 2017; 15,1607-1612.
  • El-Husseini AE, Vincent SR. Cloning and characterization of a novel RING finger protein that interacts with class V myosins. J Biol Chem. 1999; 274(28):19771-7.
  • El-Husseini AE, Kwasnicka D, Yamada T, Hirohashi S, Vincent SR. BERP, a novel ring finger protein, binds to alpha-actinin-4. Biochem Biophys Res Commun. 2000; 267(3):906-11.
  • El-Husseini AE, Fretier P, Vincent SR. Cloning and characterization of a gene (RFN22) encoding a novel brain expressed ring finger protein (BERP) that maps to human chromosome 11p15.5. Genomics. 2000; 71:363-7.
  • Mukherjee S, Tucker-Burden C, Zhang C, Moberg K, Read R, Hadjipanayis C, et al. Drosophila brat and human ortholog TRIM3 maintain stem cell equilibrium and suppress brain tumorigenesis by attenuating Notch nuclear transport, Canc. Res. 2016; 76:2443-2452.
  • Piao MY, Cao HL, He NN, Xu MQ, Dong WX, Wang WQ, et al. Potential role of TRIM3 as a novel tumour suppressor in colorectal cancer (CRC) development. Scand J Gastroenterol. 2016; 51:572-582.
  • Fu H, Yang H, Zhang X, Wang B, Mao J, LiX, et al. Exosomal TRIM3 is a novel marker and therapy target for gastric cancer. J Exp Clin Cancer Res. 2018; 37:162.
  • Song Y, Guo Q, Gao S, Hua K. Tripartite motif-containing protein 3 plays a role of tumor inhibitor in cervical cancer. Biochem Biophys Res Commun. 2018; 498: 686-692.
  • Chen G, Kong J, Tucker-Burden C, Anand M, Rong Y, Rahman F, et al. Human Brat ortholog TRIM3 is a tumor suppressor that regulates asymmetric cell division in glioblastoma. Cancer Res. 2014; 74(16): 4536-4548.
  • Liu Y, Raheja R, Yeh N, Ciznadija D, Pedraza AM, Ozawa T, et al. TRIM3, a tumor suppressor linked to regulation of p21 Waf1/Cip1. Oncogene. 2014; 33:308-15.
  • Li Y, Zhu H, Wang J, Qian X, Li N. miR-4513 promotes breast cancer progression through targeting TRIM3. Am J Transl Res. 2019; 11(4):2431-2438.
  • Wang X, Zhang Y, Pei X, Guo G, Xue B, Duan X, et al. TRIM3 inhibits P53 signaling in breast cancer cells. Cancer Cell Int. 2020; 20:559-570.
  • Ye R, AiErken N, Kuang X, Zeng H, Shao N, Lin Y, et al. Tripartite motif-containing 3 (TRIM3) enhances ER signaling and confers tamoxifen resistance in breast cancer. Oncogenesis. 2021; 10:60.
  • Lu Q, Zhang Y, Ma L, Li D, Li M, Liu P, et al. TRIM3 Negatively Regulates Autophagy Through Promoting Degradation of Beclin1 in Ewing Sarcoma Cells. OncoTargets and Therapy. 2019; 12: 11587-11595.
  • Chao J, Zhang XF, Pan QZ, Zhao JJ, Jiang SS, Wang Y, et al. Decreased expression of TRIM3 is associated with poor prognosis in patients with primary hepatocellular carcinoma. Med Oncol. 2014; 31:102.
  • Lieber M, Mazzetta J, Nelson-Rees W, Kaplan, Todaro G. Establishment of a continuous tumor-cell line (panc-1) from a human carcinoma of the exocrine pancreas. Int J Cancer. 1975; 15:741-747.
  • Chen WH, Horoszewicz JS, Leong SS, Shimano T, Penetrante R, Sanders WH, et al. Human pancreatic adenocarcinoma: in vitro and in vivo morphology of a new tumor line established from ascites. In Vitro. 1982; 18:24-34.
  • Tan MH, Nowak NJ, Loor R, Ochi H, Sandberg AA, Lopez C, et al. Characterization of a new primary human pancreatic tumor line. Cancer Invest. 1986; 4:15-23.
  • Hatakeyama S. TRIM proteins and cancer. Nat Rev Cancer. 2011; 11(11):792-804.
  • Cambiaghi V, Giuliani V, Lombardi S, Marinelli C, Toffalorio F, Pelicci PG. TRIM proteins in cancer. Adv Exp Med Biol. 2012; 770:77-91.
  • Kawai T, Akira S. Regulation of innate immune signalling pathways by the tripartite motif (TRIM) family proteins. EMBO Mol Med. 2011; 3(9):513-527.
  • Koi M, Johnson LA, Kalikin LM, Little PF, Nakamura Y, Feinberg AP. Tumor cell growth arrest caused by subchromosomal transferable DNA fragments from chromosome 11. Science. 1993; 260:361-4.
  • Boulay JL, Stiefel U, Taylor E, Dolder B, Merlo A, Hirth F. Loss of heterozygosity of TRIM3 in malignant gliomas. BMC Cancer. 2009; 9:71.
  • Zhu J, Wu G, Ke Z, Cao L, Tang M, Li Z, et al. Targeting TRIM3 deletion-induced tumor-associated lymphangiogenesis prohibits lymphatic metastasis in esopha- geal squamous cell carcinoma. Oncogene. 2019; 38:2736-49.
  • Farhadi J, Goshayeshi L, Motavalizadehkakhky A, Mehrzad J, Mehrad-Majd H. Decreased expression of TRIM3 gene predicts a poor prognosis in gastric cancer. Journal of Gastrointestinal Cancer. doi.org/10.1007/s12029-020-00563-0.
  • Song Y, Guo Q, Gao S, Hua K. miR-454-3p promotes proliferation and induces apoptosis in human cervical cancer cells by targeting TRIM3. Biochemical and Biophysical Research Communications. 2019; 516:872-879.
  • Nakahira S, Nakamori S, Tsujie M, Takahashi Y, Okami J, Yoshioka S, et al. Involvement of ribonucleotide reductase M1 subunit overexpression in gemcitabine resistance of human pancreatic cancer. Int. J. Cancer. 2007; 120:1355-1363.

Investigation of Tripartite Motif-Containing Protein 3 (TRIM3) Gene Expression in Pancreatic Cancer Cells

Year 2022, , 505 - 509, 27.12.2022
https://doi.org/10.35440/hutfd.1121746

Abstract

Background: Pancreatic cancer is among those with the worst prognosis of all cancers. The Tripartite Motif-Containing Protein 3 (TRIM3) gene, as a tumor suppressor gene, plays a role as a tumor suppressor by controlling the proliferation, migration and invasion of cancer cells. The aim of this study was to investigate the expression of TRIM3 gene at the mRNA level in AsPC1, BxPC-3 and PANC-1 pancreatic cancer cell lines.

Materials and Methods: AsPC1, BxPC-3 and PANC-1 cell lines were cultured at 37˚C in an environment containing 5% CO2 and total RNA was isolated. TRIM3 gene mRNA expression level was analyzed by Quantitative Reverse Transcription PCR (RT-qPCR) method. Analysis of relative gene expression data was performed using the 2-∆∆CT method.

Results: The mRNA expression levels of TRIM3 were found to be very low in all three cell lines. In addition, when fold change was calculated, no difference was observed between cell lines.

Conclusions: TRIM3 gene plays a role as a tumor suppressor gene in the process of carcinogenesis and it has been shown that TRIM3 expression is decreased in cancer cells. Consistent with other cancer types in the literature, TRIM3 mRNA expression was found to be very low in pancreatic cancer cells. Since this study is the only study investigating the relationship between AsPC1, BxPC-3 and PANC-1 pancreatic cancer cell lines and TRIM3, it will shed light on future functional studies.

References

  • Siegel RL, Miller KD, Jemal A. Cancer Statistics, 2020. Ca Cancer J Clin. 2020; 70:7-30.
  • Sharma C, Eltawil KM, Renfrew PD, Walsh MJ, Molinari M. Advances in diagnosis, treatment and palliation of pancreatic carcinoma: 1990-2010. World J Gastroenterol 2011; 17(7):867-897.
  • Omura N, Li CP, Li A, Hong SM, Walter K, Jimeno A, et al. Genome-wide profiling of methylated promoters in pancreatic adenocarcinoma. Cancer Biol Ther. 2008; 7(7):1146-1156.
  • Jones S, Hruban RH, Kamiyama M, Borges M, Zhang X, Parsons DW, et al. Exomic Sequencing Identifies PALB2 as a Pancreatic Cancer Susceptibility Gene. Science. 2009; 324(5924): 217-219.
  • Klein AP, Wolpin BM, Risch HA, Stolzenberg-Solomon RZ, Mocci E, Zhang M, et al. Genome-wide meta-analysis identifies five new susceptibility loci for pancreatic cancer. Nature Communications. 2018; 9:556-566.
  • Wolpin BM, Rizzato C, Kraft P, Kooperberg C, Petersen GM, Wang Z, et al. Genome-wide association study identifies multiple susceptibility loci for pancreatic cancer. Nature Genetics. 2014; doi:10.1038/ng.3052.
  • Meroni G, Diez-Roux G. TRIM/RBCC, a novel class of ‘single protein RING finger’ E3 ubiquitin ligases. Bioessays. 2005; 27(11):1147-1157.
  • Huang XQ, Zhang XF, Xia JH, Chao J, Pan QZ, Zhao JJ, et al. Tripartite motif containing 3 (TRIM3) inhibits tumor growth and metastasis of liver cancer. Chin J Cancer 2017; 36: 77.
  • Wang M, Wu J, Guo Y, Chang X, Cheng T, The tripartite motif-containing protein 3 on the proliferation and cytokine secretion of rheumatoid arthritis fibroblast-like synoviocytes. Mol. Med. Rep. 2017; 15,1607-1612.
  • El-Husseini AE, Vincent SR. Cloning and characterization of a novel RING finger protein that interacts with class V myosins. J Biol Chem. 1999; 274(28):19771-7.
  • El-Husseini AE, Kwasnicka D, Yamada T, Hirohashi S, Vincent SR. BERP, a novel ring finger protein, binds to alpha-actinin-4. Biochem Biophys Res Commun. 2000; 267(3):906-11.
  • El-Husseini AE, Fretier P, Vincent SR. Cloning and characterization of a gene (RFN22) encoding a novel brain expressed ring finger protein (BERP) that maps to human chromosome 11p15.5. Genomics. 2000; 71:363-7.
  • Mukherjee S, Tucker-Burden C, Zhang C, Moberg K, Read R, Hadjipanayis C, et al. Drosophila brat and human ortholog TRIM3 maintain stem cell equilibrium and suppress brain tumorigenesis by attenuating Notch nuclear transport, Canc. Res. 2016; 76:2443-2452.
  • Piao MY, Cao HL, He NN, Xu MQ, Dong WX, Wang WQ, et al. Potential role of TRIM3 as a novel tumour suppressor in colorectal cancer (CRC) development. Scand J Gastroenterol. 2016; 51:572-582.
  • Fu H, Yang H, Zhang X, Wang B, Mao J, LiX, et al. Exosomal TRIM3 is a novel marker and therapy target for gastric cancer. J Exp Clin Cancer Res. 2018; 37:162.
  • Song Y, Guo Q, Gao S, Hua K. Tripartite motif-containing protein 3 plays a role of tumor inhibitor in cervical cancer. Biochem Biophys Res Commun. 2018; 498: 686-692.
  • Chen G, Kong J, Tucker-Burden C, Anand M, Rong Y, Rahman F, et al. Human Brat ortholog TRIM3 is a tumor suppressor that regulates asymmetric cell division in glioblastoma. Cancer Res. 2014; 74(16): 4536-4548.
  • Liu Y, Raheja R, Yeh N, Ciznadija D, Pedraza AM, Ozawa T, et al. TRIM3, a tumor suppressor linked to regulation of p21 Waf1/Cip1. Oncogene. 2014; 33:308-15.
  • Li Y, Zhu H, Wang J, Qian X, Li N. miR-4513 promotes breast cancer progression through targeting TRIM3. Am J Transl Res. 2019; 11(4):2431-2438.
  • Wang X, Zhang Y, Pei X, Guo G, Xue B, Duan X, et al. TRIM3 inhibits P53 signaling in breast cancer cells. Cancer Cell Int. 2020; 20:559-570.
  • Ye R, AiErken N, Kuang X, Zeng H, Shao N, Lin Y, et al. Tripartite motif-containing 3 (TRIM3) enhances ER signaling and confers tamoxifen resistance in breast cancer. Oncogenesis. 2021; 10:60.
  • Lu Q, Zhang Y, Ma L, Li D, Li M, Liu P, et al. TRIM3 Negatively Regulates Autophagy Through Promoting Degradation of Beclin1 in Ewing Sarcoma Cells. OncoTargets and Therapy. 2019; 12: 11587-11595.
  • Chao J, Zhang XF, Pan QZ, Zhao JJ, Jiang SS, Wang Y, et al. Decreased expression of TRIM3 is associated with poor prognosis in patients with primary hepatocellular carcinoma. Med Oncol. 2014; 31:102.
  • Lieber M, Mazzetta J, Nelson-Rees W, Kaplan, Todaro G. Establishment of a continuous tumor-cell line (panc-1) from a human carcinoma of the exocrine pancreas. Int J Cancer. 1975; 15:741-747.
  • Chen WH, Horoszewicz JS, Leong SS, Shimano T, Penetrante R, Sanders WH, et al. Human pancreatic adenocarcinoma: in vitro and in vivo morphology of a new tumor line established from ascites. In Vitro. 1982; 18:24-34.
  • Tan MH, Nowak NJ, Loor R, Ochi H, Sandberg AA, Lopez C, et al. Characterization of a new primary human pancreatic tumor line. Cancer Invest. 1986; 4:15-23.
  • Hatakeyama S. TRIM proteins and cancer. Nat Rev Cancer. 2011; 11(11):792-804.
  • Cambiaghi V, Giuliani V, Lombardi S, Marinelli C, Toffalorio F, Pelicci PG. TRIM proteins in cancer. Adv Exp Med Biol. 2012; 770:77-91.
  • Kawai T, Akira S. Regulation of innate immune signalling pathways by the tripartite motif (TRIM) family proteins. EMBO Mol Med. 2011; 3(9):513-527.
  • Koi M, Johnson LA, Kalikin LM, Little PF, Nakamura Y, Feinberg AP. Tumor cell growth arrest caused by subchromosomal transferable DNA fragments from chromosome 11. Science. 1993; 260:361-4.
  • Boulay JL, Stiefel U, Taylor E, Dolder B, Merlo A, Hirth F. Loss of heterozygosity of TRIM3 in malignant gliomas. BMC Cancer. 2009; 9:71.
  • Zhu J, Wu G, Ke Z, Cao L, Tang M, Li Z, et al. Targeting TRIM3 deletion-induced tumor-associated lymphangiogenesis prohibits lymphatic metastasis in esopha- geal squamous cell carcinoma. Oncogene. 2019; 38:2736-49.
  • Farhadi J, Goshayeshi L, Motavalizadehkakhky A, Mehrzad J, Mehrad-Majd H. Decreased expression of TRIM3 gene predicts a poor prognosis in gastric cancer. Journal of Gastrointestinal Cancer. doi.org/10.1007/s12029-020-00563-0.
  • Song Y, Guo Q, Gao S, Hua K. miR-454-3p promotes proliferation and induces apoptosis in human cervical cancer cells by targeting TRIM3. Biochemical and Biophysical Research Communications. 2019; 516:872-879.
  • Nakahira S, Nakamori S, Tsujie M, Takahashi Y, Okami J, Yoshioka S, et al. Involvement of ribonucleotide reductase M1 subunit overexpression in gemcitabine resistance of human pancreatic cancer. Int. J. Cancer. 2007; 120:1355-1363.
There are 35 citations in total.

Details

Primary Language Turkish
Subjects Clinical Sciences
Journal Section Research Article
Authors

Muradiye Acar 0000-0003-4357-5229

Publication Date December 27, 2022
Submission Date May 26, 2022
Acceptance Date December 1, 2022
Published in Issue Year 2022

Cite

Vancouver Acar M. Pankreas Kanseri Hücrelerinde Tripartite Motif-Containing Protein 3 (TRIM3) Gen Ekspresyonunun Araştırılması. Harran Üniversitesi Tıp Fakültesi Dergisi. 2022;19(3):505-9.

Harran Üniversitesi Tıp Fakültesi Dergisi  / Journal of Harran University Medical Faculty