Korean J Physiol Pharmacol.  2023 Jul;27(4):375-381. 10.4196/kjpp.2023.27.4.375.

Rectal cancer-derived exosomes activate the nuclear factor kappa B pathway and lung fibroblasts by delivering integrin beta-1

• Gao Q
• An K
• Gao Z
• Wang Y
• Ding C
• Niu P
• Lei F

• KMID: 2544139
• DOI: http://doi.org/10.4196/kjpp.2023.27.4.375

Abstract

Numerous studies have revealed the importance of tumor-derived exosomes in rectal cancer (RC). This study aims to explore the influence of tumor-derived exosomal integrin beta-1 (ITGB1) on lung fibroblasts in RC along with underlying mechanisms. Exosome morphology was observed using a transmission electron microscope. Protein levels of CD63, CD9, ITGB1, p-p65 and p65 were detected using Western blot. To determine ITGB1's mRNA expression, quantitative real-time polymerase chain reaction was used. Moreover, levels of interleukin (IL)-8, IL-1β, and IL-6 in cell culture supernatant were measured via commercial ELISA kits. ITGB1 expression was increased in exosomes from RC cells. The ratio of p-p65/p65 as well as levels of interleukins in lung fibroblasts was raised by exosomes derived from RC cells, while was reduced after down-regulation of exosomal ITGB1. The increased ratio of p-p65/p65 as well as levels of pro-inflammatory cytokines caused by exosomes from RC cells was reversed by the addition of nuclear factor kappa B (NF-κB) inhibitor. We concluded that the knockdown of RC cells-derived exosomal ITGB1 repressed activation of lung fibroblasts and the NF-κB pathway in vitro.

Keyword

Exosomes; Fibroblasts; Integrins; Lung; Rectal neoplasms

Figure

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  Fig. 1 ITGB1 expression is increased in exosomes from SW837 cells. (A) Transmission electron microscopy images of exosomes isolated from FHC and SW837 cells. (B) The protein expression of CD63, CD9 and ITGB1 was detected by Western blot. Values are presented as mean ± SD. ns, not significant. **p < 0.01.

• 

  Fig. 2 Exosomes derived from SW837 cells activate the NF-κB pathway and lung fibroblasts. (A) The protein expression of ITGB1 was detected by Western blot in the co-culture system of MRC5 cells and exosomes derived from FHC or SW837 cells. (B) The protein expression of p-p65 and p65 was detected by Western blot in the co-culture system of MRC5 cells and exosomes derived from FHC or SW837 cells. (C) Protein levels of IL-6, IL-8 and IL-1β were measured using ELISA in the co-culture system of MRC5 cells and exosomes derived from FHC or SW837 cells. Values are presented as mean ± SD. **p < 0.01.

• 

  Fig. 3 ITGB1 down-regulation blocks activation of the NF-κB pathway and lung fibroblasts induced by RC-derived exosomes. (A) The mRNA expression of ITGB1 was detected by qRT-PCR in SW837 cells. (B) The protein expression of ITGB1 was detected by Western blot in SW837 cells. (C) The protein expression of CD63, CD9 and ITGB1 was detected by Western blot in exosomes isolated from SW837 cells. (D) The protein expression of ITGB1 was detected by Western blot in MRC5 cells. (E) The protein expression of p-p65 and p65 was detected by Western blot in the co-culture system of exosomes isolated from SW837 cells and MRC5 cells. (F) Protein levels of IL-6, IL-8 and IL-1β were measured using ELISA in MRC5 cells. Values are presented as mean ± SD. ns, not significant. **p < 0.01, ***p < 0.001.

• 

  Fig. 4 Inhibition of NF-κB pathway blocks the activation effect of exosome ITGB1 on lung fibroblasts. (A) The protein expression of ITGB1 was detected by Western blot. (B) The protein expression of p-p65 and p65 was detected by Western blot. (C) Protein levels of IL-6, IL-8 and IL-1β were measured using ELISA. Values are presented as mean ± SD. ns, not significant. **p < 0.01, ***p < 0.001.

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