Expression profiles of LMAN2 in breast cancer cell lines in hypoxia and normoxia conditions

Öz

Abstract Purpose: Breast cancer is one of the most common and leading causes of death in women. The mechanism by which breast cancer develops is not fully understood. Understanding the mechanism of initiation and the genes and proteins involved in this process may help us to fight this type of cancer. The LMAN2 gene encodes VIP36, which transports properly folded proteins. The main aim of this study was to investigate the expression of the LMAN2 gene at the molecular level in breast cancer cells with different functional defects. Determining the level of LMAN2 gene expression under hypoxia, known as oxygen deprivation, has a significant impact on tumourigenesis and metastasis, and obtaining new data on the relationship between hypoxia and ER stress was identified as a secondary objective. Material and methods: In this study, the expression level of the LMAN2 gene will be examined in breast cancer cell lines (SKBR3, MDA-MB-231, MDA-MB-468, MCF-7) and CRL4010 cell line as a control. Results: LMAN2 gene expression level was evaluated at 48-hour periods by providing normoxic and hypoxic conditions. While the LMAN2 gene expression level in MCF-7, MDA-MB-231, and SK-BR-3 cells is significantly reduced, it was highly expressed in MDA-MB-468 in hypoxic and normoxic conditions. CHOP, HERP, and BiP gene expression levels were significantly higher in MDA-MB-468 under hypoxic conditions like LMAN2 expression. Conclusion: LMAN2 and other ER stress response elements showed different expression profiles in SK-BR-3, MDA-MB-231, MDA-MB-468, and MCF7 cell lines under hypoxic conditions. Increased expression was found in MCF-7 and MDA-MB-468 cell lines, but decreased expression was detected in SK-BR-3 and MDA-MB-231 cell lines. The underlying reason for this difference is thought to be that the cell lines have different molecular properties, such as triple negative or HER2 (+/-) status.

Anahtar Kelimeler

• ER stress
• hypoxia
• LMAN2
• breast cancer
• apoptosis

Hipoksi ve normoksi koşullarında meme kanseri hücre hatlarında LMAN2'nin ifade profilleri

Öz

Amaç: Meme kanseri kadınlarda en sık görülen ve ölüme neden olan kanser türleri arasındadır. Meme kanserinin başlama mekanizması tam olarak açık değildir. Ortaya çıkış mekanizmasını ve bu süreçte yer alan genleri ve proteinleri anlamak, bu kanser türüyle mücadelemizde yardımcı olabileceğini düşünmekteyiz. LMAN2 geni, doğru katlanmış proteinleri taşıyan VIP36'yı kodlar. Bu çalışmanın temel amacı, çeşitli fonksiyonel bozukluklara sahip meme kanseri hücrelerinde LMAN2 geninin ekspresyonunu moleküler düzeyde incelemektir. Oksijen eksikliği koşulları olarak bilinen hipoksi altında LMAN2 gen ekspresyon düzeyinin saptanması, tümörigenez ve metastaz üzerinde önemli bir etkiye sahiptir ve hipoksi ile ER stresi arasındaki ilişki hakkında yeni veriler elde edilmesi de ikincil bir hedef olarak belirlenmiştir. Gereç ve yöntem: Bu çalışmada, meme kanseri hücre hatlarında (SKBR3, MDA-MB-231, MDA-MB-468, MCF-7) ve kontrol olarak CRL4010 hücre hattında LMAN2 geninin ifade düzeyi incelendi. Bulgular: LMAN2 gen ekspresyon düzeyi normoksik ve hipoksik koşullar sağlanarak 48 saatlik periyotta değerlendirilmiştir. MCF-7, MDA-MB-231 ve SK-BR-3 hücrelerinde LMAN2 gen ekspresyon düzeyi önemli ölçüde azalırken, MDA-MB-468'de hipoksik ve normoksik koşullarda yüksek düzeyde eksprese edildi. CHOP, HERP ve BiP gen ekspresyon seviyeleri, LMAN2 ekspresyonu gibi hipoksik koşullar altında MDA-MB-468'de önemli ölçüde daha yüksekti. Sonuç: LMAN2 ve diğer ER stres yanıt elemanları hipoksik koşullar altında SK-BR-3, MDA-MB-231, MDA-MB-468 ve MCF7 hücre hatlarında farklı ekspresyon profilleri göstermiştir. MCF-7 ve MDA-MB-468 hücre hatlarında artmış ekspresyon bulunurken, SK-BR-3 ve MDA-MB-231 hücre hatlarında azalmış ekspresyon tespit edilmiştir. Bu farklılığın altında yatan nedenin, hücre hatlarının üçlü negatif veya HER2 (+/-) durumu gibi farklı moleküler özelliklere sahip olması olduğu düşünülmektedir.

Anahtar Kelimeler

• ER stres
• hipoksi
• LMAN2
• meme kanseri
• apoptoz

Kaynakça

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