Knockdown of lncRNA PCAT6 suppresses the growth of non-small cell lung cancer cells by inhibiting macrophages M2 polarization via miR-326/KLF1 axis

Bioengineered. 2022 May;13(5):12834-12846. doi: 10.1080/21655979.2022.2076388.

Abstract

Non-small cell lung cancer (NSCLC) is the most common malignant tumor of lung, which seriously threatens the life of people. It has been reported that lncRNA prostate cancer-associated transcript 6 (PCAT6) could facilitate the metastasis of NSCLC cells. However, whether lncRNA PCAT6 in NSCLC cells could affect the tumor microenvironment (TME) remains unclear. In the present study, the level of PCAT6 in NSCLC cells was detected using RT-qPCR. The effects of PCAT6 knockdown on the viability and apoptosis in NSCLC cells were detected with CCK-8 and flow cytometry assay. NSCLC cell-derived exosomes were isolated with ultracentrifugation. Next, transwell assay was conducted to assess the migration and invasion of NSCLC cells. Dual-luciferase reporter assay was performed to verify the relationship among PCAT6, miR-326, and KLF1 in A549 cells. In addition, nanoparticle tracking analysis (NTA) was applied to detect the particle size of isolated exosomes. Moreover, ELISA assay was performed to detect the levels of IL-1β and IL-10 in the supernatant of macrophage. We found knockdown of PCAT6 significantly inhibited the viability, migration, and invasion of NSCLC cells. In addition, dual-luciferase reporter assay illustrated that miR-326 was the target of PCAT6 and KLF1 was the target of miR-326 in NSCLC cells. Moreover, NSCLC cells-derived exosomes could promote macrophages M2 polarization by transporting PCAT6. Meanwhile, macrophages M2 polarization was able to promote the metastasis and epithelial-mesenchymal transition (EMT) process of NSCLC cells via regulating PCAT6/miR-326/KLF1 axis. Taken together, knockdown of lncRNA PCAT6 suppressed the growth of NSCLC cells by inhibiting macrophages M2 polarization via miR-326/KLF1 axis.

Keywords: KLF1; Non-small cell lung cancer (NSCLC); lncRNA prostate cancer-associated transcript 6 (PCAT6); macrophages M2 polarization; miR-326; tumor microenvironment (TME).

MeSH terms

  • Carcinoma, Non-Small-Cell Lung* / genetics
  • Carcinoma, Non-Small-Cell Lung* / pathology
  • Cell Proliferation / genetics
  • Humans
  • Lung Neoplasms* / genetics
  • Lung Neoplasms* / pathology
  • Macrophages / pathology
  • Male
  • MicroRNAs* / genetics
  • RNA, Long Noncoding* / genetics
  • Tumor Microenvironment / genetics

Substances

  • MIRN326 microRNA, human
  • MicroRNAs
  • RNA, Long Noncoding

Grant support

The study was supported by the role and mechanism of lysosomal pathway in docetaxel against gastric cancer; Zhejiang Provincial Natural Science Foundation of China (LQ18H160024); Honokial induces apoptosis of lung squamous cell carcinoma by targeting FGF2/FGFR1 autocrine loop; Zhejiang Provincial Natural Science Foundation of China Natural Science Foundation of Zhejiang Province (LQ19H160036).