Geri Çekildi: Küçük Hücreli Akciğer Kanseri’nde RAB27A’nın CRISPR/Cas9 Sistemi Aracılığıyla Susturulmasının Biyolojik Etkileri

Yıl 2023, Cilt: 2 Sayı: Kongre Özel Sayısı - 3. Uluslararası Multidisipliner Kanser Araştırmaları Kongresi, 112 - 116, 31.10.2023

Kubilay Incı , Büşra Çelikkaya , Nesrin İrep , Aziz Gültekin , Onur Tokgün

https://doi.org/10.59312/ebshealth.1367257

Bu makale 30 Kasım 2023 tarihinde geri çekildi. https://dergipark.org.tr/tr/pub/ebshealth/issue/81016/1394394

Öz

Small cell lung cancer (SCLC) is characterized by rapid growth and early metastasis. Identifying new molecular targets are important in the pathogenesis of SCLC in order to develop new treatment strategies. RAB27A is the critical protein for intracellular exosome trafficking and is a driver of tumour progression. However, demonstrating the potential impact of suppressing RAB27A in SCLC as therapeutic approach is an important deficiency. RAB27A gene knockout SCLC cell lines were generated using a CRISPR/cas9 system. qRT-PCR, Western blotting, and Sanger sequencing were performed to confirm RAB27A knockout in SCLC cells. TEM and EXOCET assays were used to detect the alteration of exosomes. Proliferation and colony formation were detected by MTT and microscopy. Subsequently, we intrapulmonally injected N417 and H524 SCLC cells(control and RAB27A knockout for each cell) into SCID mice. The effects of RAB27A knockout on mouse tumor model were analysed using 18F-FDG PET/CT scans. Knocking out RAB27A significantly decreased the expression of CD9, CD63, Tsg101, exosome secretion and exosomal protein in SCLC(p<0.0001). We found that RAB27A knockout dramatically reduced proliferation and colony formation in SCLC cells(p< 0.001, p<0.0001). Furthermore, RAB27A knockout decreased proliferation and especially metastasis in mouse model (p<0,0001). These studies clearly demonstrated that RAB27A plays an important role in the pathogenesis of SCLC, and targeting the RAB27A gene in SCLC cell lines significantly reduces the activity of the exosomal pathway. RAB27A, therefore, can be a promising cancer therapeutic strategy.

Anahtar Kelimeler

RAB27A, exosome, SCLC, CRISPR/Cas9, Carcinogenesis

Destekleyen Kurum

This study was supported by the TUBITAK under the project number 220S104.

Proje Numarası

220S104

Geri Çekildi: Biological Effects of CRISPR/Cas9-mediated Knockout of RAB27A in SCLC

Bu makale 30 Kasım 2023 tarihinde geri çekildi. https://dergipark.org.tr/tr/pub/ebshealth/issue/81016/1394394

Öz

Small cell lung cancer (SCLC) is characterized by rapid growth and early metastasis. Identifying new molecular targets are important in the pathogenesis of SCLC in order to develop new treatment strategies. RAB27A is the critical protein for intracellular exosome trafficking and is a driver of tumour progression. However, demonstrating the potential impact of suppressing RAB27A in SCLC as therapeutic approach is an important deficiency. RAB27A gene knockout SCLC cell lines were generated using a CRISPR/cas9 system. qRT-PCR, Western blotting and Sanger sequencing were performed to confirm RAB27A knockout in SCLC cells. TEM and EXOCET assays were used to detect the alteration of exosomes. Proliferation and colony formation were detected by MTT and microscopy. Subsequently, we intrapulmonally injected N417 and H524 SCLC cells(control and RAB27A knockout for each cell) into SCID mice. The effects of RAB27A knockout on mouse tumor model were analysed using 18F-FDG PET/CT scans.Knocking out RAB27A significantly decreased the expression of CD9, CD63, Tsg101, exosome secretion and exosomal protein in SCLC(p<0.0001). We found that RAB27A knockout dramatically reduced proliferation and colony formation in SCLC cells(p< 0.001, p<0.0001). Furthermore, RAB27A knockout decreased proliferation and especially metastasis in mouse model (p<0,0001). These studies clearly demonstrated that RAB27A plays an important role in the pathogenesis of SCLC, and targeting the RAB27A gene in SCLC cell lines significantly reduces the activity of the exosomal pathway. RAB27A, therefore, can be a promising cancer therapeutic strategy.

Anahtar Kelimeler

RAB27A, exosome, SCLC, CRISPR/Cas9, Carcinogenesis

Proje Numarası

220S104

Kaynakça

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