CNN3 acts as a potential oncogene in cervical cancer by affecting RPLP1 mRNA expression

Lili Xia,Ya-Nan Zhang,Yongfang Yue,Mingyue Li,Weiguo Lu,Lu Zhao,Xing Xie,Xinyu Wang

doi:10.1038/s41598-020-58947-y

Lili Xia, Ya-Nan Zhang + Show 6 more

Open Access

https://doi.org/10.1038/s41598-020-58947-y

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Abstract

The prognosis of advanced stage cervical cancer is poorer due to cancer invasion and metastasis. Exploring new factors and signalling pathways associated with invasiveness and metastasis would help to identify new therapeutic targets for advanced cervical cancer. We searched the cancer microarray database, Oncomine, and found elevated calponin 3 (CNN3) mRNA expression in cervical cancer tissues. QRT-PCR verified the increased CNN3 expression in cervical cancer compared to para-cancer tissues. Proliferation, migration and invasion assays showed that overexpressed CNN3 promoted the viability and motility of cervical cancer cells, the opposite was observed in CNN3-knockdown cells. In addition, xenografted tumours, established from SiHa cells with CNN3 knockdown, displayed decreased growth and metastasis in vivo. Furthermore, RNA-sequencing showed that ribosomal protein lateral stalk subunit P1 (RPLP1) was a potential downstream gene. Gene function experiments revealed that RPLP1 had the same biological effects as CNN3 did. Rescue experiments demonstrated that the phenotypes inhibited by CNN3 silencing were partly or completely reversed by RPLP1 overexpression. In conclusion, we verified that CNN3 acts as an oncogene to promote the viability and motility of cervical cancer cells in vitro and accelerate the growth and metastasis of xenografted tumours in vivo, by affecting RPLP1 expression.

Highlights

Cervical cancer is the fourth most common malignancy and the fourth leading cause of cancer death in women worldwide, with an estimated 570,000 new cases and 311,000 deaths globally in 20181
By analysis of an mRNA dataset named Pyeon Multi-cancer from the Oncomine database, we found that calponin 3 (CNN3) mRNA expression was higher in 20 cervical cancer samples than in 8 normal cervix samples (Fig. 1A)
Considering that HPV oncoproteins act as drivers in the initiation and development of cervical cancer, we examined the effect of HPV16E5/E6/E7 on CNN3 expression in cervical cancer cell lines, SiHa and CaSki, and found decreased expression of CNN3 protein in both cells when they were transfected with E5 and E6/E7 siRNA, respectively (Fig. 1D)

Summary

Introduction

Cervical cancer is the fourth most common malignancy and the fourth leading cause of cancer death in women worldwide, with an estimated 570,000 new cases and 311,000 deaths globally in 20181. Due to unpopularization of screening in developing countries many patients are diagnosed at the advanced disease stage, with metastases of lymph nodes and other organs at their initial visit. Invasiveness and metastasis are the main features and cause of death in cancers with advanced stages. Finding new targets to block cancer invasion and metastasis would help improve the prognosis of advanced cervical cancer patients. We verified the CNN3 overexpression in cervical cancer tissues, and its role in promoting proliferation, migration, and invasion in cervical cancer cells and accelerating the growth and metastasis of xenografted tumours in immunodeficient mice. We identified ribosomal protein lateral stalk subunit P1, known as RPLP1, is controlled transcriptionally by CNN3, and participates in the CNN3-mediated regulation of the viability and motility of cervical cancer cells. Our findings provide evidence that CNN3 acts as an oncogene and may serve as a potential target for blocking the metastasis of cervical cancer

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