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CUL4A silencing attenuates cervical carcinogenesis and improves Cisplatin sensitivity

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Abstract

CUL4A is an ubiquitin ligase deregulated in numerous pathologies including cancer and even hijacked by viruses for facilitating their survival and propagation. However, its role in Human papilloma virus (HPV)-mediated cervical carcinogenesis remains elusive. The UALCAN and GEPIA datasets were analyzed to ascertain the transcript levels of CUL4A in cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) patients. Subsequently, various biochemical assays were employed to explore the functional contribution of CUL4A in cervical carcinogenesis and to shed some light on its involvement in Cisplatin resistance in cervical cancer. Our UALCAN and GEPIA datasets analyses reveal elevated CUL4A transcript levels in cervical squamous cell carcinoma and endocervical adenocarcinoma (CESC) patients that correlate with adverse clinicopathological parameters such as tumor stage and lymph node metastasis. Kaplan–Meier plot and GEPIA assessment depict poor prognosis of CESC patients having high CUL4A expression. Varied biochemical assays illustrate that CUL4A inhibition severely curtails hallmark malignant properties such as cellular proliferation, migration, and invasion of cervical cancer cells. We also show that CUL4A knockdown in HeLa cells causes increased susceptibility and better apoptotic induction toward Cisplatin, a mainstay drug used in cervical cancer treatment. More interestingly, we find reversion of Cisplatin-resistant phenotype of HeLa cells and an augmented cytotoxicity towards the platinum compound upon CUL4A downregulation. Taken together, our study underscores CUL4A as a cervical cancer oncogene and illustrates its potential as a prognosis indicator. Our investigation provides a novel avenue in improving current anti-cervical cancer therapy and overcoming the bottle-neck of Cisplatin resistance.

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Data availability

All data generated or analyzed in this work are included within the main article.

Abbreviations

TCGA:

The Cancer Genome Atlas

GEPIA:

Gene Expression Profiling Interactive Analysis

HPV:

Human papillomavirus

DNA:

Deoxyribonucleic acid

CUL4A:

Cullin4A

UV:

Ultraviolet

HIV:

Human immunodeficiency virus

HBV:

Hepatitis B virus

EBV:

Epstein–barr virus

SV5:

Simian virus 5

HPVI2:

Human para influenza virus type 2

EMT:

Epithelial-to-mesenchymal transition

PARP:

Poly (ADP-ribose) polymerase

pATM:

Phosphorylated Ataxia-telangiectasia mutated

pChk1:

Phosphorylated Checkpoint kinase 1

DSB:

Double-strand break

CDK:

Cyclin-dependent kinase

CDDP:

Cis-diamminedichloroplatinum(II)

MTT:

(3-(4,5-Dimethylthiazol-2-yl)-2,5-Diphenyltetrazolium Bromide)

DMSO:

Dimethyl sulfoxide

HCT-116:

Human colorectal carcinoma cells

NER:

Nucleotide excision repair

HeLa-CR:

HeLa-Cisplatin-resistant cells

IAP:

Inhibitors of apoptosis proteins

O.D.:

Optical density

FACS:

Fluorescence activated cell sorting

EDTA:

Ethylenediamine tetraacetic acid

PBS:

Phosphate buffer saline

γ-H2AX:

Phosphorylated Histone2A

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Acknowledgements

We thank CIF, UDSC, for providing FACS facility

Funding

This work was supported by Faculty Research Programme Grant from Institution of Eminence, University of Delhi (lOE/FRP/LS/2020/27), DU-DST (PURSE Grant: RC/2014/7114), UGC-SAP Grant (F.3.3./2016 DRS II UGC-SAP II), CSIR EMR Grant (37(1682)/17/EMR-II), and DBT Grant (BT/PR15422/MED/30/1705/2016) to AN. Financial assistance from SERB/JC Bose program Ref# SR/S2/JCB-08/2010 to DPS is duly acknowledged. Fellowships were provided by UGC to YA, CSIR to HB, and DBT to NS.

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Authors and Affiliations

  1. Department of Biochemistry, University of Delhi South Campus, Benito Juarez Road, New Delhi, 110021, India

    Yama Atri, Hina Bharti, Nandini Sahani, Debi P. Sarkar & Alo Nag

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  1. Yama Atri
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  2. Hina Bharti
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  3. Nandini Sahani
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  4. Debi P. Sarkar
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  5. Alo Nag
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Contributions

YA contributed to conceptualization, methodology, data curation, validation, investigation, formal analysis, writing: original draft, and visualization. HB contributed to methodology, data curation, formal analysis, writing: review and editing, and visualization. NS contributed to data curation during revision, data analysis, writing: review. DPS contributed to writing: review and editing and funding acquisition. AN conceptualization, methodology, resources, writing: review and editing, supervision, project administration, and funding acquisition. All the authors read and approved the final version of the manuscript.

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Correspondence to Alo Nag.

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Atri, Y., Bharti, H., Sahani, N. et al. CUL4A silencing attenuates cervical carcinogenesis and improves Cisplatin sensitivity. Mol Cell Biochem 479, 1041–1058 (2024). https://doi.org/10.1007/s11010-023-04776-2

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