Determination of Insulin Receptor Substrate 1 (irs1) Induced Mirna Expression Profile in Breast Cancer Cell Line
Abstract
Meme kanseri, moleküler seviyede heterojen bir yapı gösteren ve dünya çapında kadınlar arasında en sık görülen kanser türüdür. İnsülin reseptörü substratı (IRS) proteinleri, insülin veya insülin benzeri büyüme faktörü reseptörlerinden aldığı sinyalleri hücre içi ortama ileten sitoplazmik adaptör moleküllerdir. IRS1 ekspresyonundaki artışların, meme kanserinin gelişiminde ve ilerlemesinde rol oynadığı iyi bilinmektedir. IRS1 proteinlerinin ekspresyonlarına ek olarak, meme kanserinin gelişiminde ve ilerlemesinde miRNA'ların da rol aldığı gösterilmiştir. MiRNA'lar, tipik olarak mRNA'ların 3′UTR'si ile eşleşerek hedef genlerini baskılayan endojen küçük (19–25 baz) kodlamayan tek sarmallı RNA nükleotidleridir. Genellikle onkogenik veya tümör baskılayıcı olarak görev yapan miRNA'ların düzensiz ekspresyon profillerinin, meme kanserinde hücre proliferasyonu, tümörigenez ve metastaz gibi çeşitli hücresel süreçlerde yer aldığı gösterilmiştir. Bu çalışmada, meme kanseri hücre hattında insülin reseptörü substratı 1 (IRS1) indüklü miRNA ekspresyon profilinin belirlenmesi amaçlandı. Çalışmamızda, insan IRS1 geni ile transfekte edilmiş CRL4010 hücrelerinde ve insan IRS1 geni transfekte edilmemiş CRL4010 hücrelerinde (kontrol), insan IRS1 geni ile transfekte edilmiş MCF7 hücrelerinde ve insan IRS1 geni transfekte edilmemiş MCF7 hücrelerinde 47 miRNA'nın ekspresyon profilinin belirlenmesi hedeflenmiştir. Ektopik IRS1 ekspresyon seviyeleri, western blot yöntemiyle doğrulandı. Elde ettiğimiz sonuçlara göre MCF7 (kontrol) hücrelerine kıyasla ektopik IRS1 ekspresyonlu MCF7 hücrelerinde miR-181d, miR-210, miR-661, miR-130a, miR-191, miR-212, miR-93, miR-193b, miR-214, miR-21, miR-96, miR-221, miR-222, miR-373 ve miR-155 miRNA'ların ekspresyonlarının anlamlı seviyelerde arttığı belirlendi. Sonuç olarak, ektopik IRS1 ekspresyonunun meme kanseri gelişimi ve ilerlemesinde, 15 miRNA'nın ekspresyon profillerinin düzenlenmesinde rol oynadığını gözlemledik. Bu aday miRNA'ların meme kanseri gelişimindeki fonksiyonel rollerinin açıklanması için ileri çalışmalar yapılmalıdır. Anahtar Kelimeler: Meme kanseri, İnsülin sinyal yolağı, IRS1, MikroRNA profilleme
Breast cancer is the most common type of cancer among women worldwide, with a heterogeneous structure at the molecular level. Insulin receptor substrate (IRS) proteins are the cytoplasmic adapter proteins that transmit signals from insulin or insulin-like growth factor receptors to the intracellular environment. It is well established that overexpression of IRS1 plays a role in the development and progression of breast cancer. In addition to the expression of IRS1 proteins, miRNAs have also been shown to play a role in the development and progression of breast cancer. MiRNAs are endogenous small (19–25 bases) non-coding single-stranded RNA nucleotides that typically pair with the 3′UTR of mRNAs to repress their target genes. Abnormal expression profiles of miRNAs, which generally act as oncogenic or tumor suppressors, have been shown to be involved in various cellular processes such as cell proliferation, tumorigenesis and metastasis in breast cancer. In this study, it was aimed to determine the insulin receptor substrate 1 (IRS1) induced miRNA expression profile in breast cancer cell line. In our study, we tried to figure out the expression profile of 47 miRNAs in CRL4010 cells transfected with human IRS1 gene and untranfected CRL4010 cells used as control, MCF7 cells transfected with human IRS1 gene and untransfected MCF7 cells used as control. Ectopic IRS1 expression levels were confirmed by western blot method. Our results showed that expressions of miR-181d, miR-210, miR-661, miR-130a, miR-191, miR-212, miR-93, miR-193b, miR-214, miR-21, miR-96, miR-221, miR-222, miR-373 and miR-155 significantly increased in IRS1 overexpressed MCF7 cells compared to untransfected MCF7 cells. In conclusion, we observed that ectopic IRS1 expression is involved in the regulation of expression profiles of 15 miRNAs in breast cancer development and progression. Further studies should be performed to clarify the exact functional roles of these candidate miRNAs in breast cancer development. Keywords: Breast cancer, Insulin signaling pathway, IRS1, MicroRNA profiling
Breast cancer is the most common type of cancer among women worldwide, with a heterogeneous structure at the molecular level. Insulin receptor substrate (IRS) proteins are the cytoplasmic adapter proteins that transmit signals from insulin or insulin-like growth factor receptors to the intracellular environment. It is well established that overexpression of IRS1 plays a role in the development and progression of breast cancer. In addition to the expression of IRS1 proteins, miRNAs have also been shown to play a role in the development and progression of breast cancer. MiRNAs are endogenous small (19–25 bases) non-coding single-stranded RNA nucleotides that typically pair with the 3′UTR of mRNAs to repress their target genes. Abnormal expression profiles of miRNAs, which generally act as oncogenic or tumor suppressors, have been shown to be involved in various cellular processes such as cell proliferation, tumorigenesis and metastasis in breast cancer. In this study, it was aimed to determine the insulin receptor substrate 1 (IRS1) induced miRNA expression profile in breast cancer cell line. In our study, we tried to figure out the expression profile of 47 miRNAs in CRL4010 cells transfected with human IRS1 gene and untranfected CRL4010 cells used as control, MCF7 cells transfected with human IRS1 gene and untransfected MCF7 cells used as control. Ectopic IRS1 expression levels were confirmed by western blot method. Our results showed that expressions of miR-181d, miR-210, miR-661, miR-130a, miR-191, miR-212, miR-93, miR-193b, miR-214, miR-21, miR-96, miR-221, miR-222, miR-373 and miR-155 significantly increased in IRS1 overexpressed MCF7 cells compared to untransfected MCF7 cells. In conclusion, we observed that ectopic IRS1 expression is involved in the regulation of expression profiles of 15 miRNAs in breast cancer development and progression. Further studies should be performed to clarify the exact functional roles of these candidate miRNAs in breast cancer development. Keywords: Breast cancer, Insulin signaling pathway, IRS1, MicroRNA profiling
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Tıbbi Biyoloji, Gen ifadesi, Hücre dizisi, Meme hastalıkları, Meme neoplazmları, Mikro RNA, Neoplazmlar, RNA, İnsülin reseptör substratı, Medical Biology, Gene expression, Cell line, Breast diseases, Breast neoplasms, Micro RNA, Neoplasms, RNA, Insulin receptor substrate
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