MicroRNA-381 targets G protein-Coupled receptor 34 (GPR34) to regulate the growth, migration and invasion of human cervical cancer cells

https://doi.org/10.1016/j.etap.2020.103514Get rights and content

Highlights

  • MicroRNA-381 is downregulated in human cervical cancer.

  • MicroRNA-381 acts as tumor-suppressor in cervical cancer cells.

  • MicroRNA-381 inhibits the migration and invasion of cervical cancer cells.

  • MicroRNA-381 exerts its effects by targeting G protein-Coupled receptor 34.

Abstract

MicroRNAs (miRNAs) have emerged as the vital post-transcriptional regulators and control the growth and progression of different cancers types. The current study aimed at exploration of the role of microRNA-381 (miRNA-381) in human cervical cancer with emphasis on the evaluation of the underlying molecular mechanism. The results revealed a significant (P < 0.05) downregulation of miRNA-381 was found in cervical cancer tissues and cancer cell lines. Overexpression of miRNA-381 in cervical cancer cells significantly (P < 0.05) inhibited their proliferation through the induction of cell apoptosis which was accompanied by depletion of Bcl-2 and increase in Bax expression. Additionally, the cleavage of caspase-3 and 9 was also activated upon miRNA-381 overexpression. The Overexpression of miRNA-381 further inhibited the migration and invasion of cervical cancer cells. In silico analysis together with dual luciferase assay revealed G protein-Coupled receptor 34 (GPR34) to be the target of miRNA-381. The expression of GPR34 was significantly (P < 0.05) upregulated in the cervical cancer tissues and cell lines. Nonetheless, miRNA-381 overexpression caused a remarkable decrease in the expression of GPR34. The GPR34 knockdown and overexpression proved that the tumor-suppressive effects of miRNA-381 are mediated via GPR34. The study elucidated the essence of miRNA-381/GPR34 molecular regulatory axis in cervical cancer and unraveled the possibility of targeting this molecular axis as an important therapeutic approach against human cervical cancer.

Introduction

Cervical cancer is a worldwide public health issue accounting for approximately 300,000 deaths annually (Cohen et al., 2019). In 2012, cervical cancer was the 4th most prevalent disease in women and the 7th around the world, representing approximately 9 of every 10 deaths in less developed regions (Small et al., 2017; Shrestha et al., 2018). The currently employed treatment procedures against the human cervical cancer include the surgical excision of the affected tissue followed by standard chemotherapeutic approaches (Li et al., 2016a,b). Despite of fair progress in the treatment measures of cervical cancer, the crucial treatment drawbacks including poor prognosis and recurrence of the disease pose major impediments in achieving the ideal treatment success. Therefore, it becomes very crucial to evaluate the mechanics of cervical cancer at the molecular level so as to better devise the strategies of cervical cancer treatment. The micro RNAs (miRNAs) are short but conserved class of non-coding class of eukaryotic RNAs which aid in the regulation of gene expression at post-transcriptional level (Gulyaeva and Kushlinskiy, 2016). miRNAs perform the silencing of protein coding genes by binding to their untranslated regions (UTRs) at the translational level (Duchaine and Fabian, 2019). Researchers have shown that miRNAs play key regulatory role in growth and progression of human cancers (Vannini et al., 2018). Micro RNA-381 (miRNA-381) is seen to occupy the chromosomal region, 14q32.31 (Formosa et al., 2014). The miRNAs belonging to this cluster have been shown to affect the cellular behavior and influence tumor development (Zehavi et al., 2012). Previously, the miRNA-381 was shown to target transmembrane protein 16A (TMEM16A) to regulate the proliferation and metastasis of gastric cancer cells (Cao et al., 2017). Besides, the downregulation of miRNA-381 has been shown to be associated with human osteosarcoma highlighting its prognostic potential (Li et al., 2016a,b). In a recent study, a long non-coding RNA DLEU1 was found to exert tumor-suppressive effects by interacting with miRNA-381 in cervical cancer (Liu et al., 2018). Nonetheless, the role of miRNA-381 in cervical cancer yet largely unclear. Therefore, the current study explored the role and therapeutic implications of miRNA-381 in human cervical cancer via modulation of GPR34 expression.

Section snippets

Procurement of tissue specimens, culturing of cell lines and transfection of cancer cells

The procurement of 42 tissue samples pertaining to cervical cancer and normal-surrounding areas was made from the cancer patients admitted in Luoyang Central Hospital Affiliated to Zhengzhou University, Luoyang, Henan, China, prior to the application of chemo or radio-therapies. The characteristics of the cervical cancer patient donors are listed in Table 1. The patients were in-advance informed and duly signed consent was taken from them and the collection and usage of clinical specimens were

Downregulation of miRNA-381 in cervical cancer

To verify whether dysregulation of miRNA-381 accompanies the human cervical cancer, the transcriptional expression of miRNA-381 was examined in clinical tissues pertaining to cervical cancer and normal surrounding areas. It was seen that cancerous tissues exhibit significantly lower (upto 12-fold) miRNA-381 expression (Fig. 1A). When the expression study was performed in the cervical cancer cell lines (DoTc2, siHa, C33-A, CaSki and HeLa) using normal cervical epithelial cell line (NCEC) as

Discussion

The G-protein coupled receptors (GPCRs) comprise the largest family of the signal-mediating receptors (Liebmann and Bohmer, 2000). The GPCRs operate at the surface of the eukaryotic cells and influence a large number of biological and physiological cellular functions. Growing evidences suggest that these receptors are involved in the progression and metastasis of human cancers (Li et al., 2005). Besides, the GPCRs have been reported to regulate the cancer cell survival and angiogenesis of human

Conclusion

The results of present study established the prognostic role of miRNA-381 in cervical cancer. Moreover, the study demonstrated the importance of miRNA-381/GPR34 molecular regulatory axis in cervical cancer. The results indicated that diminishing transcript levels of miRNA-381 in cervical cancer relieve GPR34 from post-transcriptional silencing which accounts for growth and progression of cervical cancer. The current study thus unveils the possibility of utilizing the targeting of

Conflict of interest

The authors declare no conflict of interest.

Declaration of Competing Interest

The authors report no declarations of interest.

References (27)

  • R.T. Dorsam et al.

    G-protein-coupled receptors and cancer

    Nat. Rev. Cancer

    (2007)
  • T.F. Duchaine et al.

    Mechanistic insights into microRNA-mediated gene silencing

    Cold Spring Harb. Perspect. Biol.

    (2019)
  • A. Formosa et al.

    MicroRNAs, miR-154, miR-299-5p, miR-376a, miR-376c, miR-377, miR-381, miR-487b, miR-485-3p, miR-495 and miR-654-3p, mapped to the 14q32. 31 locus, regulate proliferation, apoptosis, migration and invasion in metastatic prostate cancer cells

    Oncogene

    (2014)
  • Cited by (11)

    • Advances in epigenetic modifications and cervical cancer research

      2023, Biochimica et Biophysica Acta - Reviews on Cancer
    • Cryo-EM structures of human GPR34 enable the identification of selective antagonists

      2023, Proceedings of the National Academy of Sciences of the United States of America
    View all citing articles on Scopus
    View full text