Investigation of the Immunoexpression of SVIP and UPR Pathway Proteins in Ovarian Adenocarcinoma Cell Line OVCAR-3

Öz

Ovarian cancer is the deadliest gynecological cancer. The endoplasmic reticulum (ER), a vital cell organelle, is involved in the folding, synthesis, and modification of a wide range of soluble and insoluble proteins. ER stress initiates the unfolded protein response (UPR), an evolutionary conserved cell stress mechanism. The UPR is mediated by three ER transmembrane sensors: IRE1, ATF6, and PERK. An inhibitor of ERAD is a small VCP/p97-interacting protein (SVIP). The study aimed to investigate the relationship between SVIP and the ER stress protein markers in the human ovarian cancer cell line OVCAR-3. The SVIP and GRP78, PERK, ATF4 immunoexpression levels were analyzed. Furthermore, employing immunofluorescence, the colocalization of three ER sensors and SVIP was ascertained. The immunoexpression of SVIP and GRP78, ATF4, and PERK were shown in the OVCAR-3 cell line. Additionally, immunofluorescence results showed the colocalization of SVIP and UPR-related proteins in the cytoplasm of OVCAR-3 cells. In conclusion, we demonstrated the cellular localization of SVIP and the proteins involved in the UPR pathway. However, further studies are needed to determine the relation between SVIP and these proteins in cancer cells.

Anahtar Kelimeler

• Endoplasmic reticulum (ER) stress
• Immunofluorescence
• Ovarian adenocarcinoma cell line (OVCAR-3)
• Small VCP-interacting protein (SVIP)
• Unfolded protein response (UPR)

Ovcar-3 Ovarian Adenokarsinoma Hücre Hattında SVIP ve UPR Proteinlerinin İmmünoekspresyonunun Araştırılması

Öz

En ölümcül jinekolojik kanser yumurtalık kanseridir. Hayati bir hücre organeli olan endoplazmik retikulum (ER), çok çeşitli çözünür ve çözünmez proteinlerin katlanması, sentezi ve modifikasyonunda rol oynar. ER stresi, evrimsel süreçte korunmuş bir hücre stres mekanizması olan katlanmamış protein yanıtını (UPR) başlatır. UPR’a, üç ER transmembran sensörü aracılık eder: IRE1, ATF6 ve PERK. ERAD inhibitörü küçük VCP/p97 ile etkileşen proteindir (SVIP). Çalışmanın amacı, Ovcar-3 insan ovarian adenokarsinoma hücre hattında SVIP ile ER stres protein belirteçleri arasındaki ilişkiyi araştırmaktır. SVIP ve GRP78, PERK, ATF4 immünoekspresyon düzeyleri analiz edildi. Ayrıca, immünofloresan kullanılarak, üç ER sensörünün ve SVIP'in kolokalizasyonu belirlendi. SVIP ve GRP78, ATF4 ve PERK'nin immünoekspresyonu OVCAR-3 hücre hattında gösterildi. Ek olarak, immünofloresan sonuçları OVCAR-3 hücrelerinin sitoplazmasında SVIP ve UPR ile ilişkili proteinlerin kolokalizasyonunu gösterdi. Sonuç olarak, SVIP'nin hücresel lokalizasyonunu ve UPR yolunda yer alan proteinleri gösterdik ancak kanser hücrelerinde SVIP ile bu proteinler arasındaki ilişkiyi belirlemek için daha fazla çalışmaya ihtiyaç vardır.

Anahtar Kelimeler

• Endoplazmik retikulum (ER) stresi
• İmmünofloresan
• Ovarian adenokarsinoma hücre hattı (OVCAR-3)
• Küçük VCP ile etkileşen protein (SVIP)
• Katlanmamış protein yanıtı (UPR)

Kaynakça

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