Detection of HPV and Human Chromosome Sites by Dual-Color Fluorescence In Situ Hybridization Reveals Recurrent HPV Integration Sites and Heterogeneity in Cervical Cancer

Human papillomavirus (HPV) integration in the human genome is suggested to be an important cause of cervical cancer. With the development of sequencing technologies, an increasing number of integration “hotspots” have been identified. However, this HPV integration information was derived from analysis of whole cervical cancer tissue, and we know very little about the integration in different cancer cell subgroups or individual cancer cells. This study optimized the preparation of probes and provided a dual-color fluorescence in situ hybridization (FISH) method to detect HPV integration sites in paraffin-embedded cervical cancer samples. We used both HPV probes and site-specific probes: 3p14 (FHIT), 8q24 (MYC), 13q22 (KLF5/KLF12), 3q28 (TP63), and 5p15 (TERT). We detected HPV signals in 75 of the 96 cases of cervical cancer; 62 cases showed punctate signals, and 13 cases showed diffuse punctate signals. We identified 3p14 as a high-frequency HPV integration site in 4 cervical cancer cases. HPV integration at 8p14 occurred in 2 cases of cervical cancer. In the same cervical cancer tissue of sample No.1321, two distinct subgroups of cells were observed based on the HPV probe but showed no difference in cell and nucleus morphology. Our study provides a new method to investigate the frequent HPV integration sites in cervical cancer and reports the heterogeneity within cervical cancer from the perspective of HPV integration.

The E545 Mutation of PIK3CA in Cervical Cancer Promotes the Survival by Several Different Pathways

Backgroud: The E545 mutation of PIK3CA in Cervical cancer is frequently happened. But the role of E545 mutation of PIK3CA in Cervical cancer is not clear.Methods: In this study, we analysised the molecular signatures of E545 mutation Cervical cancer by bioinformatics methods.Results: We collected transcriptome sequencing results of 227 no mutation cervical cancer tissue samples and 36 mutation cervical cancer tissue samples, then analyzed the data combining bioinformatics methods. A total of 5 differential expression miRNAs were obtained, including 3 up-regulated miRNAs, 1 down-rugulated miRNA. A total of 174 differential expression genes were obtained, including 132 up-regulated genes, 40 down-rugulated genes. GO analysis suggested that the up-regulated DEGs were mainly enriched in transcription factor activity, leukotriene signaling pathway and so on. Besides, we constructed a PPI network with DEGs to screen the top hub genes with a relatively high degree of connectivity. Among them CAV1, KRT20, FOS, had a degree of connectivity larger than 5 and functioned as hub module genes to promote the survival of E545 mutation cervical cancer. We also identified different miRNA-DEG axis, including hsa-mir-449a-AXL, hsa-mir-508-CGA, COL15A1, NNMT, hsa-mir-552-CHST6, NWD1. These axis regulated the survival of E545 mutation cervical cancer togetherly. Conclusions: In conclusion, this study identified DEGs and screened the key genes and pathways closely related to E545 mutation in Cervical cancer by bioinformatics analysis, These results might hold promise for finding potential therapeutic targets of cervical cancer harboring E545 mutation of PI3KCA.

The deubiquitinase (DUB) USP13 promotes Mcl-1 stabilisation in cervical cancer

Protein ubiquitination is a critical regulator of cellular homeostasis. Aberrations in the addition or removal of ubiquitin can result in the development of cancer and key components of the ubiquitination machinery serve as oncogenes or tumour suppressors. An emerging target in the development of cancer therapeutics are the deubiquitinase (DUB) enzymes that remove ubiquitin from protein substrates. Whether this class of enzyme plays a role in cervical cancer has not been fully explored. By interrogating the cervical cancer data from the TCGA consortium, we noted that the DUB USP13 is amplified in ~15% of cervical cancer cases. We confirmed that USP13 expression was increased in cervical cancer cell lines, cytology samples from patients with cervical disease and in cervical cancer tissue. Depletion of USP13 inhibited cervical cancer cell proliferation. Mechanistically, USP13 bound to, deubiquitinated and stabilised Mcl-1, a pivotal member of the anti-apoptotic BCL-2 family. Furthermore, reduced Mcl-1 expression partially contributed to the observed proliferative defect in USP13 depleted cells. Importantly, the expression of USP13 and Mcl-1 proteins correlated in cervical cancer tissue. Finally, we demonstrated that depletion of USP13 expression or inhibition of USP13 enzymatic activity increased the sensitivity of cervical cancer cells to the BH3 mimetic inhibitor ABT-263. Together, our data demonstrates that USP13 is a potential oncogene in cervical cancer that functions to stabilise the pro-survival protein Mcl-1, offering a potential therapeutic target for these cancers.

PROFILE OF HISTOPATHOLOGY OF CERVICAL CANCER TISSUES IN PATIENTS OF THE DR PIRNGADI MEDAN HOSPITAL

Cervical cancer is a malignant neoplasm that happened in the cervical area. Several types of cervical cancer that usually happens in women such as squamous cell carcinoma (SCC), adenocarcinoma, adenosquamous carcinoma and neuroendocrine carcinoma. Therefore, this study aims to determine a descriptive description of the histopathological profile of cervical cancer tissue from patients that registered in the Anatomical Pathology Laboratory at the DR. Pirngadi Medan Hospital, in the period of 2019. The type of our research is retrospective descriptive with the sampling technique, namely total sampling. The sample used was cervical cancer tissue obtained from the biopsy and surgery on patients who were registered from January 2019 to December 2019, totaling 18 samples. Based on the descriptive histopathological description, we found three types of cervical cancer tissue: squamous cell carcinoma (55.5%), adenocarcinoma (38.8%) and adenosquamous carcinoma (5.5%). The neuroendocrine carcinoma type did not find in this present study. Of three types of these cervical cancer, we got that cervical cancer patients are women aged between 36-72 years, with an average age of 54 years.

PIM-1 may function as an oncogene in cervical cancer via activating the EGFR signaling

Background: This work was designed to explore the roles of PIM-1 in the development of cervical cancer. Methods: There were 90 paired cervical tumor samples and the non-tumor adjacent tissue. The levels of PIM-1 in different samples were examined using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) methods. The potential diagnostic value of PIM-1 was analyzed by the receiver operating characteristic (ROC) curve; furthermore, the expression of EGFR in tumor samples was detected, and Pearson’s correlation analysis was performed to analyze the relationship between the expression of PIM-1 and EGFR. Finally, cervical cancer cell line Hela cells were cultured and treated by PIM-1 siRNA, and MTT assay and Pi/Annexin V assay were performed to explore the effects of PIM-1 siRNA on the growth and apoptosis ability of the Hela cells. Results: PIM-1 was significantly up-regulated in cervical cancer tissue compared to adjacent tissue, and the expression of PIM-1 in patients with cervical cancer is positively associated with the size and metastasis of the tumor. ROC analysis showed PIM-1 is a sensitive biomarker for the diagnosis of cervical cancer. Furthermore, EGFR was over-expressed in cervical cancer tumor tissues, and the levels of PIM-1 and EGFR in cervical cancer tissue were positively correlated. Finally, PIM-1 siRNA dramatically inhibited the viability and promoted the apoptosis of the Hela cells. Conclusion: Our findings prove that PIM-1 may function as an oncogene in cervical cancer and can regulate the EGFR signaling in cervical cancer.