Cancer-Derived Exosomal miR-651 as a Diagnostic Marker Restrains Cisplatin Resistance and Directly Targets ATG3 for Cervical Cancer

Objective. Cancer-derived exosomes can facilitate drug resistance in cervical cancer. However, the mechanisms remain elusive. Herein, we observed the roles of exosomal miR-651 in cisplatin resistance of cervical cancer. Methods. Circulating miR-651 was detected in cervical cancer and healthy individuals. The diagnostic efficacy was determined. When transfected with miR-651 mimics, cisplatin resistance, apoptosis, and proliferation were assessed. The cancer-derived exosomes were separated and identified. We observed the uptake of PKH67-labeled exosomes by HeLa/S cells. After coculture with exosomes secreted by HeLa/S or HeLa/DDP cells, malignant behaviors were examined in HeLa/S cells. The interactions between ATG3 and miR-651 were validated by dual luciferase report. Biological behaviors were investigated for HeLa/S cells cocultured with exosomes secreted by miR-651 mimic-transfected HeLa/DDP cells. Results. Downregulated circulating miR-651 was found in cancer subjects than healthy individuals. It possessed high sensitivity and accuracy in diagnosing cervical cancer ( AUC = 0.9050 ). Lower miR-651 expression was confirmed in HeLa/DDP than HeLa/S cells. Forced miR-651 lessened cisplatin resistance and proliferation and elevated apoptosis in HeLa cells. ATG3 was a direct target of miR-651. The exosomes isolated from HeLa cells were rich in CD63, CD9, and CD81 proteins, thereby identifying the isolated exosomes. Exosomes secreted by HeLa/DDP cells can be absorbed by HeLa/S cells. When being cocultured with exosomes secreted by HeLa/DDP cells, malignant behaviors of HeLa/S cells were enhanced, which were ameliorated by miR-651 mimic exosomes. Conclusion. Our findings showed that cancer-derived exosomal miR-651 restrained cisplatin resistance and directly targeted ATG3, indicating that exosomal miR-651 could be a therapeutic agent.

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Nucleolin Promotes Cisplatin Resistance in Cervical Cancer by the YB1-MDR1 Pathway

Journal of Oncology ◽

10.1155/2021/9992218 ◽

2021 ◽

Vol 2021 ◽

pp. 1-11

Author(s):

Jing Ke ◽

Chunming Gu ◽

Heyan Zhang ◽

Yang Liu ◽

Wenhao Zhang ◽

...

Keyword(s):

Cervical Cancer ◽

Drug Resistance ◽

Hela Cells ◽

Protein Level ◽

Molecular Mechanisms ◽

Cisplatin Resistance ◽

Mdr1 Gene ◽

Luciferase Reporter ◽

Drug Efflux ◽

Cell Line Hela

Purpose. Cervical cancer is the fourth most common cancer in women worldwide and is the main cause of cancer-related deaths in women. Cisplatin (DDP) is one of the major chemotherapeutic drugs for cervical cancer patients. But, drug resistance limits the effectiveness of cancer therapy. Nucleolin (NCL) is a nucleocytoplasmic multifunctional protein involved in the development of cancer. It has been reported that NCL may be a potential target for modulation of drug resistance. However, the precise molecular mechanisms are poorly understood. Materials and Methods. Human cervical cancer Hela cells and their cisplatin-resistant cell line Hela/DDP were used in this study. The protein level of NCL in cervical cancer cells was measured by western blot analysis. Hela cells and Hela/DDP cells were transfected with NCL overexpression plasmid or NCL siRNA separately. MTT and EdU assay were performed to evaluate the cell viability and sensitivity to cisplatin. The drug efflux function of MDR1 protein was assessed by intracellular rhodamine-123 accumulation assay.The promoter activity of MDR1 was assessed by using a dual-luciferase reporter assay. Results. We found that the protein level of NCL was elevated in Hela/DDP cells. Overexpression of NCL increased cervical cancer cell proliferation and attenuated the sensitivity to cisplatin. Overexpression of NCL increased Multidrug resistance (MDR1) gene expression and drug efflux. Our results demonstrated that NCL was highly related with cisplatin resistance in cervical cancer. NCL played an important role in MDR1 gene transcription through regulation of the transcription factor YB1. Conclusion. Our findings revealed the novel role of NCL in cisplatin-resistant cervical cancer and NCL may be a potential therapeutic target for chemoresistance.

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Recent advances in the contribution of noncoding RNAs to cisplatin resistance in cervical cancer

PeerJ ◽

10.7717/peerj.9234 ◽

2020 ◽

Vol 8 ◽

pp. e9234

Author(s):

Xin Wen ◽

Shui Liu ◽

Jiyao Sheng ◽

Manhua Cui

Keyword(s):

Cervical Cancer ◽

Drug Resistance ◽

Cancer Cells ◽

Treatment Option ◽

Disease Burden ◽

Cisplatin Resistance ◽

Noncoding Rnas ◽

Mechanisms Of Action ◽

Female Population ◽

Cellular Processes

Cervical cancer (CC) remains a major disease burden on the female population worldwide. Chemotherapy with cisplatin (cis-diamminedichloroplatinum (II); CDDP) and related drugs are the main treatment option for CC; however, their efficacy is limited by the development of drug resistance. Noncoding RNAs (ncRNAs) have been found to play critical roles in numerous physiological and pathological cellular processes, including drug resistance of cancer cells. In this review, we describe some of the ncRNAs, including miRNAs, lncRNAs and circRNAs, that are involved in the sensitivity/resistance of CC to CDDP-based chemotherapy and discuss their mechanisms of action. We also describe some ncRNAs that could be therapeutic targets to improve the sensitivity of CC to CDDP-based chemotherapy.

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Usefulness of SS18-SSX antibody as a diagnostic marker for pulmonary metastatic synovial sarcoma

10.21203/rs.3.rs-386235/v1 ◽

2021 ◽

Author(s):

Kentaro Miura ◽

Kimihiro Shimizu ◽

Takashi Eguchi ◽

Sachie Koike ◽

Shunichiro Matsuoka ◽

...

Keyword(s):

Synovial Sarcoma ◽

Specific Antibody ◽

Diagnostic Marker ◽

High Sensitivity ◽

University Hospital ◽

The Novel ◽

Diagnostic Efficacy ◽

Lung Cancers ◽

Touch Imprint Cytology

Abstract Background The novel SS18-SSX fusion-specific antibody is reported to have high sensitivity and specificity for the diagnosis of primary synovial sarcoma (SS), which often metastasizes to the lung. Thus far, no study has validated the diagnostic efficacy of SS18-SSX antibody for pulmonary metastatic SS, and this is the first study to report these findings. We aimed to investigate the usefulness of the SS18-SSX antibody in the diagnosis of pulmonary metastatic SS. Methods We evaluated the immunohistochemistry of SS18-SSX fusion-specific antibody (E9X9V) in 10 pulmonary metastatic SS cases and the corresponding five primary sites (four limbs and one mediastinum) in five patients (SS diagnosis of was already confirmed by fluorescence in-situ hybridization in the metastatic and primary sites), and in 93 clinical and histologic mimics including 49 non-SS, pulmonary metastatic sarcomas, 39 primary lung cancers, and five intrathoracic solitary fibrotic tumors. All specimens were surgically resected at Shinshu University Hospital during 2001–2019. For primary and metastatic SS, we also evaluated SS18-SSX immunohistochemistry in needle biopsy and touch imprint cytology specimens from the primary site. Results SS18-SSX had stained diffusely strong in all 10 pulmonary metastatic SS cases and the corresponding five primary sites without staining of SS18-SSX in all 93 clinical and histologic mimics (100% sensitivity and 100% specificity). SS18-SSX had sufficiently stained in the biopsy and cytology specimens. Conclusions Immunohistochemistry of the SS18-SSX fusion-specific antibody is useful for the differential diagnosis of pulmonary metastatic SS in clinical practice. This simple and reliable method can replace traditional genomic tests.

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Effect of miR-301a/PTEN pathway on the proliferation and apoptosis of cervical cancer

Innate Immunity ◽

10.1177/1753425919840702 ◽

2019 ◽

Vol 25 (4) ◽

pp. 217-223 ◽

Cited By ~ 3

Author(s):

Li-na Peng ◽

Wen-tian Shi ◽

Huan-rong Feng ◽

Chuan-yu Wei ◽

Qi-nan Yin

Keyword(s):

Cervical Cancer ◽

Cervical Carcinoma ◽

Hela Cells ◽

Pten Expression ◽

Cell Counting ◽

Luciferase Assay ◽

Loss Of Function ◽

Reverse Pattern ◽

Significant Difference ◽

Direct Target

The aim of this study was to evaluate the effect of the miR-301a/PTEN pathway in cervical cancer. miR-301a and PTEN expression were measured by quantitative real-time PCR (qRT-PCR) in tissues samples and HeLa cells. PTEN protein level was determined by Western blotting. Dual reporter luciferase assay was performed to validate PTEN as a direct target of miR-301a. The gain- and loss-of function assay was performed by miR-301a overexpression and silencing. Cell proliferation was monitored by cell counting Kit-8 (CCK-8). Cell apoptosis was quantitated by flow cytometry. SPSS was used to analyze the significant difference in the treatments. miR-301a demonstrated a significantly higher expression in cervical carcinoma tissues compared with the paired non-carcinoma tissues ( n = 12), while PTEN expression was found to be significantly lower in cervical carcinoma tissues than their paired non-carcinoma tissues ( n = 12). In addition, PTEN was identified as the direct target of miR-301a. Moreover, overexpression of miR-301a significantly promoted HeLa cells proliferation and anti-apoptosis which had a reverse pattern after PTEN overexpression. Our results confirm PTEN as a direct target of miR-301a in HeLa cells and suggest that miR-301a/PTEN pathway contributes to the development and progression of cervical cancer.

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miR-145 Contributes to the Progression of Cervical Carcinoma by Directly Regulating FSCN1

Cell Transplantation ◽

10.1177/0963689719861063 ◽

2019 ◽

Vol 28 (9-10) ◽

pp. 1299-1305 ◽

Cited By ~ 7

Author(s):

Li Ma ◽

Ling-Ling Li

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Cervical Carcinoma ◽

Hela Cells ◽

Underlying Mechanism ◽

Luciferase Assay ◽

Cancer Cell Proliferation ◽

Expression Levels ◽

Direct Target

The purpose of our study was to investigate the underlying mechanism and functional role of microRNA-145 (miR-145) in cervical cancer. In this study, quantitative real-time PCR (qRT-PCR) was used to detect miR-145 and FSCN1 expression levels in tissues and HeLa cells. Western blotting was performed to determine the protein level of FSCN1. The luciferase assay was used to verify the direct target of miR-145. The CCK-8 assay and 2D colony formation assays were performed to determine the effects of miR-145 mimics or FSCN1 silencing on cell proliferation. miR-145 expression levels were significantly down-regulated, while FSCN1 expression levels were significantly up-regulated in the cervical carcinoma tissues compared with their matched non-cancerous tissues. In addition, FSCN1 expression levels were negatively correlated to miR-145 in tissues. Next, FSCN1 was verified as the direct target of miR-145 in HeLa cells. Moreover, overexpression of miR-145 dramatically inhibited the proliferation of HeLa cells. The silencing of FSCN1 exhibited the similar patterns on cell proliferation as miR-145 overexpression. The miR-145/ FSCN1 axis contributes to the progression of cervical cancer by inhibition of cervical cancer cell proliferation.

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Usefulness of SS18-SSX antibody as a diagnostic marker for pulmonary metastatic synovial sarcoma

Diagnostic Pathology ◽

10.1186/s13000-021-01110-6 ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Kentaro Miura ◽

Kimihiro Shimizu ◽

Takashi Eguchi ◽

Sachie Koike ◽

Shunichiro Matsuoka ◽

...

Keyword(s):

Synovial Sarcoma ◽

Specific Antibody ◽

Diagnostic Marker ◽

High Sensitivity ◽

University Hospital ◽

The Novel ◽

Diagnostic Efficacy ◽

Lung Cancers ◽

Touch Imprint Cytology

Abstract Background The novel SS18-SSX fusion-specific antibody is reported to have high sensitivity and specificity for the diagnosis of primary synovial sarcoma (SS), which often metastasizes to the lung. Thus far, no study has validated the diagnostic efficacy of SS18-SSX antibody for pulmonary metastatic SS. Therefore, we aimed to investigate the usefulness of the SS18-SSX antibody in the diagnosis of pulmonary metastatic SS. Methods We evaluated the immunohistochemistry of SS18-SSX fusion-specific antibody (E9X9V) in 10 pulmonary metastatic SS cases and the corresponding five primary sites (four limbs and one mediastinum) in five patients, for whom SS was already diagnosed and confirmed by fluorescence in-situ hybridization in the metastatic and primary sites, and in 93 clinical and histologic mimics including 49 non-SS, pulmonary metastatic sarcomas, 39 primary lung cancers, and five intrathoracic solitary fibrotic tumors. All specimens were surgically resected at Shinshu University Hospital during 2001–2019. For primary and metastatic SS, we also evaluated SS18-SSX immunohistochemistry using needle biopsy and touch imprint cytology specimens from the primary site. Results SS18-SSX staining was diffusely-strongly positive in all 10 pulmonary metastatic SS cases and the corresponding five primary sites; whereas, it was negative in all 93 clinical and histologic mimics (100% sensitivity and 100% specificity). Further, SS18-SSX staining was also sufficiently positive in the biopsy and cytology specimens. Conclusions Immunohistochemistry of the SS18-SSX fusion-specific antibody is useful for the differential diagnosis of pulmonary metastatic SS in clinical practice. This simple and reliable method has the potential to replace traditional genomic tests. However, further studies are warranted in this regard.

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Sema4C mediates EMT inducing chemotherapeutic resistance of miR-31-3p in cervical cancer cells

Scientific Reports ◽

10.1038/s41598-019-54177-z ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 6

Author(s):

Li Jing ◽

Wang Bo ◽

Feng Yourong ◽

Wang Tian ◽

Wang Shixuan ◽

...

Keyword(s):

Cervical Cancer ◽

Drug Resistance ◽

Cancer Cells ◽

Cisplatin Resistance ◽

Malignant Tumors ◽

Chemotherapeutic Resistance ◽

Novel Approach ◽

Cervical Cancer Cells ◽

Cancer Tissues

AbstractSema4C, the target of many miRNAs, is involved in EMT-mediated chemotherapeutic resistance of many malignant tumors. However, the underlying upstream regulatory mechanisms of Sema4C-induced EMT and Sema4C-mediated drug resistance are still unclear. The aim of this study was to explore the potential role of miR-31-3p/Sema4C in regulating EMT in cisplatin-resistant (CR) cervical cancer cells. High expression levels of Sema4C were more frequently found in cervical cancer tissues and were associated with poor prognosis, whereas miR-31-3p was significantly downregulated in cervical cancer tissues, which was associated with shorter disease-free and overall survival. Overexpression of miR-31-3p inhibited malignant behaviors and EMT of cervical cancer cells in vitro. Furthermore, miR-31-3p was identified to directly target Sema4C, and upregulation of miR-31-3p reversed EMT-mediated biological functions, including cisplatin resistance of Sema4C in cervical cancer cells. These results suggest that Sema4C promoted EMT-mediated cisplatin resistance in cervical cancer cells and that this effect was inhibited by overexpression of miR-31-3p. Thus, silencing Sema4C or overexpression of miR-31-3p could be a novel approach to treat drug resistance to chemotherapy in cervical cancers.

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Effects of Papaya Leaf Extract (Carica papaya L.) on Cellular Proliferation and Apoptosis in Cervical Cancer Mice Model

Phytothérapie ◽

10.3166/phyto-2018-0096 ◽

2019 ◽

Vol 17 (5) ◽

pp. 265-275

Author(s):

Y. Peristiowati ◽

Y. Puspitasari ◽

Indasah

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Hela Cells ◽

Leaf Extract ◽

Caspase 3 ◽

Methanol Extract ◽

Negative Control ◽

Proliferation Index ◽

Control Group ◽

P53 Expression

This study is aimed at analyzing the anticancer properties of papaya leaf extract, specifically the inhibition of cell proliferation and apoptotic induction through nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) and p53 pathways. Twenty-five mice (Mus musculus), aged 2 months and weighing 20–30 g, was injected with 0.5 mg dexamethasone for 7 days. The mice were then injected intracutaneously with 1 ml of HeLa cells (8 × 106 HeLa cells/microliter). The mice were divided into five groups (5 each): negative control (P1) (5% CMC-Na, sodium carboxymethyl cellulose), treatment II (225 mg/kg BW (body weight) papaya leaves methanol extract), treatment III (450 mg/kg BW), treatment IV (750 mg/kg BW), and treatment PV (2 mg alcohol anticancer drug). Papaya leaf extract treatments were applied for 2 weeks. Then, the tumor tissue was isolated for hematoxylin and eosin staining. Immunohistochemical imaging was used to detect Ki-67, caspase-3, NF-κB, and p53 expression. Further analysis was undertaken using the ImmunoRatio software program. The results indicated that administration of papaya leaf methanol extract significantly increased the expression of NF-κB and p53 at a dose of 450 mg/kg BW. Our results also showed that the mice treated with 450 mg of papaya leaf extract per kg of BW (P3) had the largest increase of caspase-3 expression compared to the negative control group. Papaya leaf ethanol extract decreased the cancer cell proliferation index and increased apoptosis of cancer cells in animal models of cervical cancer; it may also work to increase NF-kB expression and expression of the p53 gene.

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E6 and E7 oncoproteins: Potential targets of cervical cancer

Current Medicinal Chemistry ◽

10.2174/0929867327666201111145546 ◽

2020 ◽

Vol 27 ◽

Author(s):

Ramarao Malla ◽

Mohammad Amjad Kamal

Keyword(s):

Cervical Cancer ◽

Drug Resistance ◽

Host Cell ◽

Molecular Mechanisms ◽

Immune Therapy ◽

Cervical Lesions ◽

Diagnostic Strategies ◽

E6 And E7 Oncoproteins ◽

E6 And E7

: Cervical cancer (CC) is the fourth leading cancer in women in the age group 15-44 globally. Experimental as well as epidemiological studies identified that type16 and 18 HPV cause 70% of precancerous cervical lesions as well as cervical cancer worldwide by bringing about genetic as well as epigenetic changes in the host genome. The insertion of the HPV genome triggers various defense mechanisms including the silencing of tumor suppressor genes as well as activation of oncogenes associated with cancer metastatic pathway. E6 and E7 are small oncoproteins consisting of 150 and 100 amino acids respectively. These oncoproteins affect the regulation of the host cell cycle by interfering with p53 and pRb. Further these oncoproteins adversely affect the normal functions of the host cell by binding to their signaling proteins. Recent studies demonstrated that E6 and E7 oncoproteins are potential targets for CC. Therefore, this review discusses the role of E6 and E7 oncoproteins in metastasis and drug resistance as well as their regulation, early oncogene mediated signaling pathways. This review also uncovers the recent updates on molecular mechanisms of E6 and E7 mediated phytotherapy, gene therapy, immune therapy, and vaccine strategies as well as diagnosis through precision testing. Therefore, understanding the potential role of E6/E7 in metastasis and drug resistance along with targeted treatment, vaccine, and precision diagnostic strategies could be useful for the prevention and treatment of cervical cancer.

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The Antitumor Efficiency of Zinc Finger Nuclease Combined with Cisplatin and Trichostatin A in Cervical Cancer Cells

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520620666200804102300 ◽

2020 ◽

Vol 20 (17) ◽

pp. 2125-2135

Author(s):

Ci Ren ◽

Chun Gao ◽

Xiaomin Li ◽

Jinfeng Xiong ◽

Hui Shen ◽

...

Keyword(s):

Cervical Cancer ◽

Cancer Cells ◽

Zinc Finger ◽

Hela Cells ◽

Colony Formation ◽

Trichostatin A ◽

Combined Therapy ◽

Hpv E7 ◽

Anti Cancer ◽

Cervical Cancer Cells

Background: Persistent infection with the high-risk of human papillomavirus (HR-HPVs) is the primary etiological factor of cervical cancer; HR-HPVs express oncoproteins E6 and E7, both of which play key roles in the progression of cervical carcinogenesis. Zinc Finger Nucleases (ZFNs) targeting HPV E7 induce specific shear of the E7 gene, weakening the malignant biological effects, hence showing great potential for clinical transformation. Objective: Our aim was to develop a new comprehensive therapy for better clinical application of ZFNs. We here explored the anti-cancer efficiency of HPV targeted ZFNs combined with a platinum-based antineoplastic drug Cisplatin (DDP) and an HDAC inhibitor Trichostatin A (TSA). Methods: SiHa and HeLa cells were exposed to different concentrations of DDP and TSA; the appropriate concentrations for the following experiments were screened according to cell apoptosis. Then cells were grouped for combined or separate treatments; apoptosis, cell viability and proliferation ability were measured by flow cytometry detection, CCK-8 assays and colony formation assays. The xenograft experiments were also performed to determine the anti-cancer effects of the combined therapy. In addition, the HPV E7 and RB1 expressions were measured by western blot analysis. Results: Results showed that the combined therapy induced about two times more apoptosis than that of ZFNs alone in SiHa and HeLa cells, and much more inhibition of cell viability than either of the separate treatment. The colony formation ability was inhibited more than 80% by the co-treatment, the protein expression of HPV16/18E7 was down regulated and that of RB1 was elevated. In addition, the xenografts experiment showed a synergistic effect between DDP and TSA together with ZFNs. Conclusion: Our results demonstrated that ZFNs combined with DDP or TSA functioned effectively in cervical cancer cells, and it provided novel ideas for the prevention and treatment of HPV-related cervical malignancies.

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