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MiRNA-211 triggers an autophagy-dependent apoptosis in cervical cancer cells: regulation of Bcl-2

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Abstract

Cervical cancer is a significant cause of morbidity and mortality in gynecological malignancies. Although autophagy plays a critical role in affecting cell apoptosis and proliferation, the role of hsa-miR-211-5p (miR-211) in modulating autophagy of cervical cancer cells remains unclear. In the current study, the level of miR-211 was downregulated in cervical cancer specimens, compared to the paired para-carcinoma tissues. While Bcl-2 was upregulated, LC3-II/I was decreased in the tumors, indicating inhibited apoptosis and autophagy. The forced expression of miR-211 inhibited proliferation, and promoted apoptosis in SiHa cervical cancer cells, evidenced by increased expression of apoptotic proteins, caspase-3, and PARP. While the miR-211 inhibitor exerted reverse effects on C-33A cervical cancer cells. Further, miR-211 induced autophagy in cervical cancer cells, as manifested by the presence of LC3 puncta, increased LC3-II/I and Beclin1 levels, and decreased p62 level. The miR-211-induced apoptosis was alleviated by an autophagy inhibitor 3-methyladenine (3-MA). In addition, Bcl-2 was identified as a target of miR-211. Besides, the apoptosis and autophagy triggered by miR-211 were attenuated by Bcl-2 in SiHa cells. In summary, our work indicates that miR-211 induced autophagy and autophagy-dependent apoptosis by regulating Bcl-2 in cervical cancer cells, which provided further understanding of autophagy in cervical carcinogenesis.

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Abbreviations

ATGs:

Autophagy-regulated genes

Atg14:

Autophagy-related gene 14

BAK:

Bcl-2 antagonist/killer

BAX:

Bcl-2-associated X protein

CCK-8:

Cell counting kit-8

CDK6:

Cyclin-dependent kinase 6

DMEM:

Dulbecco’s modified Eagle’s medium

ECL:

Enhanced chemiluminescence

EMT:

Epithelial-mesenchymal transition

FBS:

Fetal bovine serum

HPV:

Human papillomavirus

IgG-HRP:

Peroxidase-conjugated immunoglobulin G

miRNAs:

MicroRNAs

miR-211:

hsa-miR-211-5p

MEM:

Minimum essential medium Eagle

MITF:

mIcrophthalmia-associated transcription factor

MUC4:

Mucin 4

PE:

Phosphatidylethanolamine

PI:

Propidium iodide

PMSF:

Phenylmethanesulfonyl fluoride

PTEN:

Phosphatase and tensin homolog

PVDF:

Polyvinylidene fluoride

RB1CC1:

RB1 inducible coiled-coil 1

RCC:

Renal cell carcinoma

RPMI-1640:

Roswell park memorial institute-1640

SPARC:

Secreted protein acidic and rich in cysteine

SD:

Standard deviation

SDS-PAGE:

Sodium dodecylsulphate polyacrylamide gel electrophoresis

SOX4:

Sex-determining region Y-related high mobility group box 4

ZEB2:

Zinc finger E-box binding homeobox 2

3′UTR:

3′untranslated region

3-MA:

3-methyladenine

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Funding

This study was supported by grants from the National Natural Science Foundation of China (No. 81602272), the Natural Science Foundation of Heilongjiang Province (No. H2017012), and the Startup Foundation for Post-doctor of Heilongjiang Province (No. LBH-Q16145).

Author information

Shang Liu and Liyuan Guo conceived and designed this study. Shang Liu, Hongyan Wang, Jing Mu, Hao Wang, Yan Peng, Qi Li, and Dongwei Mao carried out the experiments, data analysis, and results interpretation. Shang Liu, Hongyan Wang, Jing Mu, and Liyuan Guo performed supervision and manuscript writing. All authors contributed to final approval of the version.

Correspondence to Liyuan Guo.

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All procedures were conducted according to Declaration of Helsinki, and approved by Ethic Committee of Cancer Hospital Affiliated of Harbin Medical University.

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The authors declare that they have no conflict of interest.

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Liu, S., Wang, H., Mu, J. et al. MiRNA-211 triggers an autophagy-dependent apoptosis in cervical cancer cells: regulation of Bcl-2. Naunyn-Schmiedeberg's Arch Pharmacol 393, 359–370 (2020). https://doi.org/10.1007/s00210-019-01720-4

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Keywords

  • Cervical cancer
  • miR-211
  • Bcl-2
  • Apoptosis
  • Autophagy