Silencing LncRNA-NFATC3 Enhances The Radiosensitivity Of Cervical Cancer HeLa Cells

Background:Cervical cancer is a common malignancy affecting women,posing a serious threat to their health.While radiotherapy remains a primary treatment option for cervical cancer and offers significant clinical benefits for patients,tolerance to radiotherapy remains a major contributor to treatment failure.DNA damage repair is a key factor in decreasing radiotherapy sensitivity in cervical cancer patients.Therefore,investigating the mechanisms behind radiotherapy sensitivity in cervical cancer is of significant clinical importance and practical value.Long non-coding RNAs(LncRNAs)are non-protein coding RNA molecules that are over 200 nucleotides in length and play a vital role in regulating the biological behavior of cancer.Recent studies have shown that certain LncRNAs play a crucial role in regulating sensitivity to radiotherapy in cervical cancer,though the specific molecular mechanisms remain unclear.Our group’s previous sequencing data revealed that LncRNA-NFATC3 expression was significantly increased in tissues resistant to cervical cancer radiotherapy,with upregulation of m RNA related to DNA damage repair.Therefore,this study aims to further explore the role of LncRNA-NFATC3 in the biological behavior of cervical cancer tumors and its specific mechanism in regulating radiotherapy sensitivity.These results will enhance our understanding of the role of LncRNA in radiotherapy for cervical cancer and provide novel insights and strategies for the treatment of this disease.Objective:The objective of this study is to investigate the role of LncRNA-NFATC3 in regulating the biological characteristics and radiosensitivity of cervical cancer cells treated with radiation.Additionally,the study aims to elucidate the mechanism by which LncRNA-NFATC3 influences the radiosensitivity of cervical cancer by affecting the DNA damage repair pathway.Method:1.RNA-seq was conducted on LncRNA and m RNA in tissues of five patients with radiotherapy tolerance for cervical cancer before and after radiotherapy.GO enrichment analysis was conducted on the differentially expressed m RNA to identify their distribution in tumor-related signaling pathways.The results of RNA-seq were verified by q RT-PCR to ensure the reliability of the sequencing results.2.A cell model of LncRNA-NFATC3 knockout was established by constructing and transfecting si-RNA targeting LncRNA-NFATC3 into cervical cancer He La cells.3.The effects of LncRNA-NFATC3 gene expression knockdown on proliferat-ion,migration,invasion,and apoptosis of cervical cancer cells before and after radiotherapy were detected using CCK-8 assay,clonogenc assay,Transwell assay,flow cytometry,and Western blot.4.The foci of cervical cancer cell DNA marker 53BP1 before and after radiotherapy were determined by cell immunofluorescence.Results:1.RNA-seq analysis revealed that the expression of LncRNA-NFATC3 was significantly up-regulated in radiotherapy-resistant tissues of cervical cancer.This finding was further confirmed by q RT-PCR detection.GO enrichment analysis showed that the differentially expressed m RNAs were primarily involved in DNA replication,cell cycle,and DNA damage pathways.The up-regulated m RNAs detected by sequencing were also validated by q RT-PCR.2.Silencing LncRNA-NFATC3 resulted in a significant reduction in the proliferation,migration,and invasion of He La cells after radiotherapy.Furthermore,it also promoted cell apoptosis.In addition,the silencing of LncRNA-NFATC3 increased the number of damage points of 53BP1 in He La cells after radiotherapy.Conclusion:1.Our findings suggest that the inhibition of LncRNA-NFATC3 can effectively suppress the proliferation,invasion,and metastasis of cervical cancer cells under irradiation.Moreover,it can also promote cell apoptosis and DNA damage,leading to an increase in the sensitivity of cervical cancer cells to radiotherapy.2.DNA damage related proteins may be involved in the regulation radiotherapy sensitivity of cervical cancer,but its specific regulatory mechanism and whether it is correlated with LncRNA-NFATC3 still need to be verified.These results provide a potential therapeutic target for enhancing the efficacy of radiotherapy in cervical cancer treatment.