Long non-coding RNA GAS5 regulates the growth and metastasis of human cervical cancer cells via induction of apoptosis and cell cycle arrest

https://doi.org/10.1016/j.abb.2020.108320Get rights and content

Abstract

Studies have proved the role of GAS5 in the development of different cancers. This study was undertaken to investigate the role and explore therapeutic implications of GAS5 in human cervical cancer. The results showed that GAS5 was significantly (p < 0.05) downregulated in human cervical cancer tissues. The results also showed that cervical cancer progresses with the suppression of GAS5 expression levels. Additionally, the expression of GAS5 was also significantly (p < 0.05) downregulated in human cervical cancer cell lines. Nonetheless, overexpression of GAS5 caused a remarkable decrease in the proliferation of C33A and HeLa cervical cancer cells. The decrease in the proliferation rate was attributed to the induction of apoptosis of C33A and HeLa cells which was accompanied with upregulation of Bax and suppression of Bcl-2. Additionally, GAS5 overexpression also promoted the arrest of C33A and HeLa cells at the G2/M check point of cell cycle via suppression of cyclin B1 and CDK1 expression. The transwell assays showed that GAS5 overexpression significantly (p < 0.05) inhibited the migration and invasion of the C33A and HeLa cervical cancer cells. The bioinformatics analysis as well as the dual luciferase assay showed GAS5 acts as a target of miR-135a. Interestingly, the expression of miR-135a was upregulated in the human cervical cancer cells and its suppression exerted growth inhibitory effects on the C33A and HeLa cells. However, silencing of GAS5 could nullify the effects of miR-135a suppression on the proliferation of C33A and HeLa cells. Taken together, the results of this study point towards the therapeutic implications of GAS5 in the treatment of cervical cancer.

Introduction

Human cervical cancer is one of the major health issues among women worldwide [1,2]. Currently, cervical cancer is ranked as the fourth most prevalent cancer type among females [3]. Of the all cervical cancer related mortality, 85% of deaths occur in the developing countries [[4], [5], [6]]. The 5-year survival rate of metastatic cervical cancer is only 16% [7]. Additionally, the molecular mechanics leading to the development of cervical cancer is poorly understood. Over the years studies have revealed the importance of long non-coding RNAs (LncRNAs) in the development and progression of human cancers including the cervical cancer [[8], [9], [10], [11]]. The lncRNAs constitute a diverse group of RNA molecules lacking open reading frame and possessing a nucleotide length greater than 200 and reaching up to 100 kb [12,13]. The lncRNAs are mainly transcribed by RNA polymerase II and may or may not be adenylated [14]. It has been reported that a vast majority of RNA moieties in humans excluding those corresponding to mitochondrial and ribosomal RNAs possess no known functions and constitute the non-coding RNAs, including the lncRNAs [15]. However, recent investigations have provided some important insights into the role of lncRNAs and have shown that some important biological functions such as development of embryos and organogenesis are greatly influenced by lncRNAs [16,17]. LncRNAs are also involved in development of human tumors and studies suggest that aberrant expression of lncRNAs is associated with the promotion of cancer onset [18,19]. The lncRNA, GAS5 (growth arrest-specific transcript 5) has been shown to play a pivotal role in number of human cancers. The downregulation of GAS5 was found to be involved in the development of hepatocellular carcinoma and responsible for the poor cancer prognosis [20]. Further, GAS5 affected the proliferation and survival of non-small lung cancer cells [21]. In yet another study it has been found to act as a tumor suppressor in renal cell carcinoma [22]. Nonetheless, the role and therapeutic implications of lncRNA GAS5 have not been explored in the human cervical cancer. Against this backdrop, the current study was designed to investigate the role and therapeutic implications of GAS5 in regulation of human cervical cancer. The results showed that GAS5 is downregulated in human cervical cancer and it negatively regulates the proliferation and metastasis of cervical cancer cells by interacting miR-135-JAK/STAT axis. Taken together, the findings of the present study point towards the therapeutic implications of GAS5 in cervical cancer treatment.

Section snippets

Collection of clinical tissues and ethical statement

A total of 65 cervical cancer tissues and 65 normal adjacent tissues were analyzed in the study. The tissue samples were obtained after proper consent from the patients undergoing treatment at Tonji Hospital, Huazhong University of Science and Technology Wuhan, China from June 2016 to December 2018. Patients were aged 36–71 years with an average age of 46.5 years. All the cancerous tissues and adjacent normal tissues were obtained from resected tumors and adjacent non-tumor tissues. The

Cervical cancer proceeds with suppression of GAS5

In order to investigate the expression pattern of GAS5 in cervical cancer, qRT-PCR was performed. The results revealed the expression of GAS5 to be significantly lower in cervical cancer tissue (Fig. 1A). Also, the clinical cancer stages were found to proceed with decreasing GAS5 expression count (Fig. 1B). The comparison of GAS5 transcript levels between non-metastatic and meta-static cancerous tissues indicated that metastatic tissues have very low expression of GAS5 (Fig. 1C). The expression

Discussion

The studies on the role of long-non coding RNA (lncRNAs) in cancer have deduced a vital linkage between the lncRNA regulatory group and human cancer development [[23], [24], [25]]. Furthermore, there is growing evidence about the involvement of lncRNA dysregulation in cancer progression [26,27]. The efficacy of anti-cancer drugs has also been seen to be influenced by lncRNAs [28]. Researchers have also identified a number of lncRNAs regulating the growth and development of human cervical

Conclusion

To sum up, the results of this study highlight the negative regulatory role of GAS5 long no-coding RNA in cervical cancer. Results indicate the molecular therapeutic potential of GAS5 and additionally suggest the drug mediated targeting of GAS5 as an alternative mode of anti-cancer approach against the human cervical cancer. Nonetheless, in vivo studies have been required to establish GAS5 as therapeutic target for the management of cervical cancer.

Declaration of competing interest

The authors declare that there are no conflicts of interest.

Acknowledgements

We acknowledge the funding support from Central Guiding Local Science and Technology Development Special Project (No: 2018ZYYD014), and National Key Research and Development Program of China (No: 2018YFC1002801).

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