Normal Cervical Cells Research Articles

Artificial classification of cervical squamous lesions in ThinPrep cytologic tests using a deep convolutional neural network.

The diagnosis of squamous cell carcinoma requires the accurate classification of cervical squamous lesions in the ThinPrep cytologic test (TCT). It primarily relies on a pathologist's interpretation under a microscope. Deep convolutional neural networks (DCNN) have played an increasingly important role in digital pathology. However, they have not been applied to diverse datasets and externally validated. In the present study, a DCNN model based on VGG16 and an ensemble training strategy (ETS) based on 5-fold cross-validation was employed to automatically classify normal and abnormal cervical squamous cells from a multi-center dataset. First, we collected a dataset comprising 82 TCT samples from four hospitals and fine-tuned our model twice on the dataset with and without the ETS. Then, we compared the classifications obtained from the models with those provided by two skilled pathologists to discriminate the performance of the models in terms of classification accuracy and efficiency. Finally, paired sample t-tests were used to validate the consistency between the classification provided by the proposed methods and that of the pathologists. The results showed that ETS slightly, though not significantly, improved the classification accuracy compared with that of the pathologists: P0=0.387>0.05 (DCNN without ETS vs. DCNN with ETS), P1=0.771>0.05 (DCNN with ETS vs. pathologist 1), P2=0.489>0.05 (DCNN with ETS vs. pathologist 2). The DCNN model was almost 6-fold faster than that of the pathologists. The accuracy of our automated scheme was similar to that of the pathologists, but a higher efficiency in the accurate identification of cervical squamous lesions was provided by the scheme. This result allows for wider and more efficient screening and may provide a replacement for pathologists in the future. Future research should address the viability of the practical implementation of such DCNN models in the laboratory setting.

Macroscopic and microscopic analysis of the mechanical properties and adhesion force of cells using a single cell tensile test and atomic force microscopy: Remarkable differences in cell types

Many experimental techniques have been reported to provide knowledge of the mechanical behavior of cells from biomechanical viewpoints, however, it is unclear how the intercellular structural differences influence macroscopic and microscopic mechanical properties of cells. The aim of our study is to clarify the comprehensive mechanical properties and cell-substrate adhesion strength of cells, and the correlation with intracellular structure in different cell types. We developed an originally designed micro tensile tester, and performed a single cell tensile test to estimate whole cell tensile stiffness and adhesion strength of normal vascular smooth muscle cells (VSMCs) and cervical cancer HeLa cells: one half side of the specimen cell was lifted up by a glass microneedle, then stretched until the cell detached from the substrate, while force was simultaneously measured. The tensile stiffness and adhesion strength were 49 ± 10 nN/% and 870 ± 430 nN, respectively, in VSMCs (mean ± SD, n = 8), and 19 ± 17 nN/% and 320 ± 160 nN, respectively, in HeLa cells (n = 9). The difference was more definite in the surface elastic modulus map obtained by atomic force microscopy, indicating that the internal tension of the actin cytoskeleton was significantly higher in VSMCs than in HeLa cells. Structural analysis with confocal microscopy revealed that VSMCs had a significant alignment of F-actin cytoskeleton with mature focal adhesion, contrary to the randomly oriented F-actin with smaller focal adhesion of HeLa cells, indicating that structural arrangement of the actin cytoskeleton and their mechanical tension generated the differences in cell mechanical properties and adhesion forces. The finding strongly suggests that the mechanical and structural differences in each cell type are deeply involved with their physiological functions.

Highly sensitive and selective detection of cancer cell with an all-optical scheme

The mechanical properties (e.g. surface tension and bending modulus) of the cell membrane usually change when normal cells become cancerous, which opens up a way to diagnose cancer by detecting the vibration of the cell membrane. In this letter, we theoretically propose a highly sensitive method for the detection of cancer cells in terms of an all-optical domain. In the case of cervical cells, we find that cervical cancer cells have a peak shift of in the absorption spectrum compared with normal cervical cells. The results show that a peak shift can be detected in the absorption spectrum once the cells become cancerous. We expect that the proposed approach could advance the research in cancer detection.

Water soluble PEGylated phenothiazines as valuable building blocks for bio-materials.

The paper reports a series of three new PEGylated phenothiazine derivatives which keep the potential of valuable building blocks for preparing eco-materials addressed to a large realm of fields, from bio-medicine to opto-electronics. They were synthetized by connecting the hydrophilic poly(ethylene glycol) to the hydrophobic phenothiazine via an ether, ester, or amide linking group. The successful synthesis of the targeted polymers and their purity were demonstrated by NMR and FTIR spectroscopy methods. Their capacity to self-assembly in water was studied by DLS and UV-vis techniques and the particularities of the formed aggregates were investigated by fluorescence spectroscopy, SEM, AFM, POM and UV light microscopy. The biocompatibility was assessed on normal human dermal fibroblasts and human cervical cancer cells. The synthetized compounds showed the formation of luminescent aggregates and proved excellent biocompatibility on normal cells. In addition, a concentration dependent cytotoxicity against HeLa cancer cells was noticed for the PEGylated phenothiazine containing an ester unit.

MicroRNA-135a-induced formation of CD133+ subpopulation with cancer stem cell properties in cervical cancer.

Cancer stem cells (CSCs) play significant roles in tumor initiation. MicroRNA-135a (miR-135a) induced the formation of a CD133+ subpopulation from a human papillomavirus-immortalized cervical epithelial cell line. Compared with the CD133- cells, the CD133+ cells expressed higher levels of miR-135a and OCT4, exhibited significantly higher tumorsphere forming capacity and the time required for tumorsphere formation was shortened in the second generation. Serum induction suppressed the expression of CD133, OCT4 and miR-135a, but increased expression of involucrin in the miR-135a-induced CD133+ cells. The miR-135a-induced CD133+ cells were tumorigenic in a limiting dilution approach in vivo. The cells expressed significantly higher level of active β-catenin and OCT4 than the CD133- counterpart. Wnt3a enhanced the expression of OCT4 and CD133 in cervical cancer cells but failed to enhance CD133 transcription in normal cervical cells. Wnt3a stimulation also increased tumorsphere size and self-renewal of miR-135a-induced CD133+ subpopulation. Wnt/β-catenin inhibition suppressed tumorsphere formation while Wnt3a partially nullified the inhibitory effect. Taken together, miR-135a induced the formation of a subpopulation of cells with CSC properties both in vitro and in vivo and the Wnt/β-catenin signaling pathway is essential to maintain its tumorigenicity.

Effect of sodium dichloroacetate as single agent or in combination with cisplatin in normal and human cervical cancer cell lines

Purpose: To evaluate the synergistic cytotoxicity of sodium dichloroacetate (DCA) in combination with cisplatin (CIS) against human cervical cancer cell lines.
 Methods: Cervical cancer SiHa and HeLa cells and normal cells (Hek-293, Vero, peripheral blood mononuclear and human erythrocytes) were treated in vitro with DCA and CIS individually or their combination. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) method while hemolytic activity was evaluated from the released hemoglobin. Halfmaximal inhibitory concentration (IC50) of DCA or CIS was obtained.
 Results: The combination of DCA + CIS decreased the cell viability of SiHa, Hek-293, Vero, and PBMC cells, but not of Hela cells (p < 0.05). Furthermore, the individual treatments alone or in combination did not cause significant hemolysis (p < 0.05).
 Conclusion: The combination of DCA + CIS increases the damage caused by CIS alone on SiHa cells. It also decreases the cell viability of Hek-293 and Vero without affecting peripheral blood mononuclear and human erythrocyte integrity. The results suggest that the combination of DCA and CIS can induce synergistic antitumor effect in different types of cancer cell lines. However, further studies are required to determine the biological effects of the combination of DCA and CIS in vivo.
 Keywords: Cervical cancer, Sodium dichloroacetate, Cisplatin, Viability, Hemolysis

Internet of health things-driven deep learning system for detection and classification of cervical cells using transfer learning

Cervical cancer is one of the fastest growing global health problems and leading cause of mortality among women of developing countries. Automated Pap smear cell recognition and classification in early stage of cell development is crucial for effective disease diagnosis and immediate treatment. Thus, in this article, we proposed a novel internet of health things (IoHT)-driven deep learning framework for detection and classification of cervical cancer in Pap smear images using concept of transfer learning. Following transfer learning, convolutional neural network (CNN) was combined with different conventional machine learning techniques like K nearest neighbor, naive Bayes, logistic regression, random forest and support vector machines. In the proposed framework, feature extraction from cervical images is performed using pre-trained CNN models like InceptionV3, VGG19, SqueezeNet and ResNet50, which are fed into dense and flattened layer for normal and abnormal cervical cells classification. The performance of the proposed IoHT frameworks is evaluated using standard Pap smear Herlev dataset. The proposed approach was validated by analyzing precision, recall, F1-score, training–testing time and support parameters. The obtained results concluded that CNN pre-trained model ResNet50 achieved the higher classification rate of 97.89% with the involvement of random forest classifier for effective and reliable disease detection and classification. The minimum training time and testing time required to train model were 0.032 s and 0.006 s, respectively.

Effects of Galectin-1 on Biological Behavior in Cervical Cancer.

Background: We previously revealed that the expression of galectin-1 (LGALS1) was significantly reduced after neoadjuvant chemotherapy treatment in cervical cancer patients. The objective of this study is to investigate the effects of LGALS1 expression on biological behaviors of cervical cancer cells.Methods: Immunohistochemistry and immunocytochemistry were performed to detect the expression of LGALS1 in cervical cancer tissues and cells (SiHa and C33A). Western blot analysis was performed to evaluate the efficacy of lentivirus-mediated upregulation or downregulation of LGALS1 in cervical cancer cells. Cell viability and proliferation were detected by CCK-8 and BrdU assays, respectively. Annexin V-FITC/PI apoptosis detection kit was employed to measure the apoptosis of cervical cancer cells. Transwell invasion and migration assays were also conducted to explore the invasive and migratory capabilities of cervical cancer cells. The expression of apoptosis- (Bcl-2 and Bax), invasion- (MMP-2 and MMP-9), and migration-related (Fascin and Ezrin) proteins, were detected by Western blot analysis. Xenograft mouse model of cervical cancer was generated to explore whether LGALS1 overexpression could promote tumor growth in vivo.Results: LGALS1 was overexpressed in cervical cancer tissues and cell lines compared to that in normal cervical tissues and epithelium cells. Upregulation of LGALS1 significantly promoted the cell proliferation, inhibited cell apoptosis, and enhanced the migratory and invasive abilities of both SiHa and C33A cells, whereas downregulation of LGALS1 led to the opposite results. The level of Bcl-2, MMP-2, MMP-9, Fascin, and Erzin expression was significantly upregulated in cervical cancer cells with LGALS1 overexpression, while converse results were obtained in LGALS1 knockdown cancer cells. In vivo study also showed that LGALS1 overexpression facilitated tumor growth of cervical cancer cells.Conclusion: Overexpression of LGALS1 significantly promoted and enhanced the aggressive features of cervical cancer both in vitro and in vivo, which may be associated with high expression of Bcl-2, MMP-2, MMP-9, Fascin, and Erzin proteins.

Comparison of Cancer Cell Elasticity by Cell Type.

Lower cellular elasticity is a distinguishing feature of cancer cells compared with normal cells. To determine whether cellular elasticity differs based on cancer cell type, cells were selected from three different cancer types including breast, cervix, and lung. For each cancer type, one counterpart normal cell and three types of cancer cells were selected, and their elasticity was measured using atomic force microscopy (AFM). The elasticity of normal cells was in the order of MCF10A > WI-38 ≥ Ect1/E6E7 which corresponds to the counterpart normal breast, lung, and cervical cancer cells, respectively. All cancer cells exhibited lower elasticity than their counterpart normal cells. Compared with the counterpart normal cells, the difference in cellular elasticity was the greatest in cervical cancer cells, followed by lung and breast cancer cells. This result indicates lower elasticity is a unique property of cancer cells; however, the reduction in elasticity may depend on the histological origin of the cells. The F-actin cytoskeleton of cancer cells was different in structure and content from normal cells. The F-actin is mainly distributed at the periphery of cancer cells and its content was mostly lower than that seen in normal cells.

Pyrrolo and pyrrolopyrimidine sulfonamides act as cytotoxic agents in hypoxia via inhibition of transmembrane carbonic anhydrases

A series of novel sulfonamide derivatives bearing pyrrole and pyrrolopyrimidine scaffolds were synthesized and screened as carbonic anhydrase inhibitors. The inhibition activity of the synthesized compounds was evaluated against the cytosolic human carbonic anhydrase isoforms I and II and the transmembranal isoforms IX and XII. Several candidates showed potent inhibitory activity against IX and XII isoforms. Furthermore, ex vivo screening of cytotoxic selectivity and activity of the most potent derivatives were carried out against normal cells (WI38) and cervical cancer cell line (HeLa) under normal and hypoxic conditions using acetazolamide as reference drug. Compound 11b potency was nearly three folds higher in hypoxic than normoxic condition whereas that of compound 11f was nearly four folds higher in hypoxic vs. normoxic HeLa cells. All the screened derivatives exhibited less potency on normal cells (WI38). Molecular docking was carried out to discover the possible binding mode of compounds within the active site of isoform CA IX.

CircAGFG1 promotes cervical cancer progression via miR-370-3p/RAF1 signaling

BackgroundIn past decades, circular RNAs (circRNAs) have achieved increasing attention because of its regulatory role in different kinds of cancers. However, how circAGFG1 regulates cervical cancer (CC) is still largely undiscovered. This study aims to evaluate the role of a novel circRNAs and related molecular mechanism in CC cells.MethodsHigh or low level of circAGFG1 was detected in CC cells or normal cell line with qRT-PCR. The proliferative and migratory abilities of CC cells were assessed with loss-of function assays. The downstream miRNA and mRNA of circAGFG1 were searched out and proved by using bioinformatics analysis and mechanism experiments. Recue assays were designed to confirm the role of circAGFG1/miR-370-3p/RAF1 axis in CC cell activities.ResultsThe levels of circAGFG1 was abundant in CC cells in comparison with normal cervical cell End1/E6E7. The inhibitory effect of decreased circAGFG1 level on the proliferative and migratory abilities of CC cells was assessed. CircAGFG1 and miR-370-3p were localized in the cytoplasm and they can interact with each other. Moreover, miR-370-3p was downregulated in CC cells. We also determined the negative effect of miR-370-3p on RAF1. CircAGFG1 could promote RAF1 expression by absorbing miR-370-3p, thereby activating RAF/MEK/ERK pathway. circAGFG1 promoted proliferation and migration of CC cells via enhancing the activity of RAF/MEK/ERK pathway by sponging miR-370-3p and further regulating RAF1.ConclusionThe results of this study provided new evidence that circAGFG1 acted as a vital regulator in cervical cancer proliferation and migration, giving great promise to apply it as a potential biomarker for diagnosis and therapy in CC treatment.

MicroRNA-15a-5p down-regulation inhibits cervical cancer by targeting TP53INP1 in vitro.

An increasing number of reports have shown that microRNAs (miRNAs) play a vital role in the occurrence and development of cancer by acting as tumor inhibitors or oncogenes. The purpose of this research was to explore whether the expression level of microRNA-15a-5p (miR-15a-5p) was related to TP53 regulated inhibitor of apoptosis 1 (TP53INP1) in cervical cancer, and to explore the role of miR-15a-5p in cervical cancer in vitro. Human cervical cancer tissues and adjacent normal tissues were obtained from 30 cervical cancer patients. Firstly, we carried out the quantitative Real Time-PCR (qRT-PCR) assay to evaluate the level of miR-15a-5p in cervical cancer tissues and cell lines. The TargetScan and the Dual-Luciferase Reporter Assay were used to confirm the relationship between TP53INP1 and miR-15a-5p. Besides, the Cell Counting Kit-8 (CCK-8) and the flow cytometry analysis were performed to detect the effect of miR-15a-5p on cell proliferation and apoptosis in cervical cancer cells. Our results showed that the expression of miR-15a-5p was enhanced in cervical cancer tissues and cells lines. The data from the Dual-Luciferase Reporter Assay demonstrated that TP53INP1 was a direct target of miR-15a-5p. We also found that TP53INP1 was down-regulated in the cervical cancer tissues and cell lines compared with the adjacent normal tissues and normal cervical cells. Besides, the down-regulation of miR-15a-5p depressed cervical cancer cell proliferation and enhanced cell apoptosis. Our results clearly suggested that the down-regulation of TP53INP1 successfully impaired the tumor-inhibition effects of miR-15a-5p inhibitor in cervical cancer cells. Our findings indicated that miR-15a-5p functioned as a tumor-promoting gene in cervical cancer by directly targeting TP53INP1, indicating that miR-15a-5p might be a potential treatment target for cervical cancer patients.

LncRNA HAND2-AS1 inhibits cancer cell proliferation, migration and invasion by downregulating ROCK1 in HPV-positive and negative cervical squamous cell carcinoma.

Long non-coding (lnc)RNA HAND2 antisense RNA 1 (HAND2-AS1) exhibited tumor suppression activity in different types of cancer. However, its role in cervical squamous cell carcinoma (CSCC), which is frequently diagnosed among females, has not been elucidated. The current study revealed that lncRNA HAND2-AS1 was downregulated, while Rho-associated protein kinase 1 (ROCK1) was upregulated in serum of human papillomavirus (HPV)-positive and -negative patients with CSCC compared with healthy controls. Correlation analysis revealed that the expression levels of lncRNA HAND2-AS1 were negatively correlated with the expression levels of ROCK1 in HPV-positive and -negative patients with CSCC but not in healthy controls. Downregulation of lncRNA HAND2-AS1 distinguished patients with CSCC from healthy controls. Additionally, lncRNA HAND2-AS1 overexpression led to reduced expression levels of ROCK1 in HPV-positive and -negative human CSCC cell lines but not in normal cervical cell lines. ROCK1 overexpression did not significantly affect the expression of lncRNA HAND2-AS1 in all the cell lines investigated. lncRNA HAND2-AS1 overexpression inhibited, while ROCK1 overexpression promoted the proliferation, migration and invasion of HPV-positive and -negative human CSCC cell lines but not normal cervical cell lines. ROCK1 overexpression attenuated the effects of lncRNA HAND2-AS1 overexpression on cancer cell proliferation, migration and invasion. lncRNA HAND2-AS1 may inhibit cancer cell proliferation, migration and invasion by downregulating ROCK1 in HPV-positive and -negative CSCC.

A protocol for Enhanced imaging and Quantification of Cervical Cell Under Scanning electron Microscope

The application of Field Emission Scanning Electron Microscopy and Energy Dispersive X-Ray (FE-SEM/EDX) for the characterization of biological samples can produce promising results for classification purpose. The limitations of the established sample preparation technique of cervical cells for FE-SEM/EDX study that differentiate between normal and abnormal cells prompted the development of a proposed protocol for the preparation of cervical cells. The proposed protocol was conducted by a McDowell-Trump fixative prepared in 0.1M phosphate buffer without osmium tetroxide at 4°C for 2 h in the fixation process. Morphologically, the cervical cells scanned under the FE-SEM/EDX did not present blackening effects, and the structure of the cells was not broken based on the FE-SEM images. Quantitatively, the possible elemental distributions in the cells, such as carbon, nitrogen, oxygen, and sodium, are detected in samples prepared by the proposed protocol. The analysed elements were validated using the Attenuated Total Reflection and Fourier Transform Infrared (ATR/FTIR) spectroscopy. Moreover, by avoiding osmium tetroxide fixation, the time required for sample preparation decreased significantly. This sample preparation protocol can be used for normal and abnormal cervical cells in achieving better results in terms of morphological, detected elemental distribution, and rapid in time.

Design and Application of a High-Throughput, High-Content Screening System for Natural Product Inhibitors of the Human Parasite Trichomonas vaginalis.

It is estimated that Trichomonas vaginalis affects an astonishing 3.9% of the world's population, and while many of those infected are asymptomatic, progression of the disease can lead to serious health problems. Currently, the nitroimidazoles constitute the only drug class approved to treat trichomoniasis in the United States, which makes the spread of drug resistance a realistic concern. We developed a new image-based, high-throughput, and high-content assay for testing natural products (purified compounds and extracts) for antitrichomonal activity. Applying this assay system to a library of fungal natural product extracts led to the identification of three general classes of natural product inhibitors that exhibited moderate to strong activities against T.vaginalis: anthraquinones, xanthone-anthraquinone heterodimers, and decalin-linked tetramic-acid-containing metabolites. The tetramate natural products emerged as the most promising candidate molecules with pyrrolocin A (51) exhibiting potent activity against the parasite (EC50 = 60 nM), yet this metabolite showed limited toxicity to mammalian cell lines (selectivity index values of 100 and 167 versus 3T3 fibroblast and Ect1 normal cervical cells, respectively). The imaging-based assay system is a powerful tool for the bioassay-guided purification of single-component antitrichomonal biomolecules from complex natural product mixtures.

Inhibiting MNK Selectively Targets Cervical Cancer via Suppressing eIF4E-Mediated β-Catenin Activation

Targeting β-catenin has been shown to have great potential therapeutic value in cervical cancer. Because β-catenin is also essential for normal cells, strategies to specifically target cancer will require identification of druggable factors capable of distinguishing β-catenin signaling pathways between cancer and normal cells. Expression of p-eIF4E and p-β-catenin was analyzed in malignant and normal cervical tissues and cells. The effects and its underlying mechanisms of targeting MNK and eukaryotic translation initiation factor 4E (eIF4E) were determined in cervical cancer and normal cells. Inhibiting MNK/eIF4E axis selectively targets cervical cancer without affecting normal cervical cells, via suppressing eIF4E-mediated β-catenin activation. We found that eIF4E phosphorylation was upregulated in cervical cancer cells and tissues but not normal cervical counterparts, and its phosphorylation at Ser 209 activates Wnt/β-catenin signaling, promotes growth and migration in cervical cancer, in an MNK-dependent manner. MNK inhibition via genetic small interfering RNA (siRNA) knockdown or pharmacologic inhibitor effectively decreased phosphorylation of eIF4E and β-catenin, leading to reduced β-catenin activity and transcript levels of Wnt target genes in cervical cancer cells. Consistently, we found that MNK kinase inhibitor is effective in inhibiting proliferation and migration, and inducing apoptosis in cervical cancer but not normal cervical cells. The combination of MNK kinase inhibitor with paclitaxel achieved greater efficacy in cervical cancer cells than paclitaxel alone. Our work identifies MNK-eIF4E axis as a specific and critical regulator of β-catenin activity in cervical cancer but not normal cervical cells, and suggests that targeting MNK is a useful therapeutic strategy in cervical cancer.

Postbiotic metabolites produced by Lactobacillus plantarum strains exert selective cytotoxicity effects on cancer cells

BackgroundLactobacillus plantarum, a major species of Lactic Acid Bacteria (LAB), are capable of producing postbiotic metabolites (PM) with prominent probiotic effects that have been documented extensively for rats, poultry and pigs. Despite the emerging evidence of anticancer properties of LAB, very limited information is available on cytotoxic and antiproliferative activity of PM produced by L. plantarum. Therefore, the cytotoxicity of PM produced by six strains of L. plantarum on various cancer and normal cells are yet to be evaluated.MethodsPostbiotic metabolites (PM) produced by six strains of L. plantarum were determined for their antiproliferative and cytotoxic effects on normal human primary cells, breast, colorectal, cervical, liver and leukemia cancer cell lines via MTT assay, trypan blue exclusion method and BrdU assay. The toxicity of PM was determined for human and various animal red blood cells via haemolytic assay. The cytotoxicity mode was subsequently determined for selected UL4 PM on MCF-7 cells due to its pronounced cytotoxic effect by fluorescent microscopic observation using AO/PI dye reagents and flow cytometric analyses.ResultsUL4 PM exhibited the lowest IC50 value on MCF-7, RG14 PM on HT29 and RG11 and RI11 PM on HL60 cell lines, respectively from MTT assay. Moreover, all tested PM did not cause haemolysis of human, dog, rabbit and chicken red blood cells and demonstrated no cytotoxicity on normal breast MCF-10A cells and primary cultured cells including human peripheral blood mononuclear cells, mice splenocytes and thymocytes. Antiproliferation of MCF-7 and HT-29 cells was potently induced by UL4 and RG 14 PM respectively after 72 h of incubation at the concentration of 30% (v/v). Fluorescent microscopic observation and flow cytometric analyses showed that the pronounced cytotoxic effect of UL4 PM on MCF-7 cells was mediated through apoptosis.ConclusionIn conclusion, PM produced by the six strains of L. plantarum exhibited selective cytotoxic via antiproliferative effect and induction of apoptosis against malignant cancer cells in a strain-specific and cancer cell type-specific manner whilst sparing the normal cells. This reveals the vast potentials of PM from L. plantarum as functional supplement and as an adjunctive treatment for cancer.

Biological Characteristics of Cervical Precancerous Cell Proliferation.

High-grade squamous intraepithelial lesions (HSILs) are regarded as precancerous lesions that can progress to cervical carcinoma; however, it is very difficult to effectively differentiate these precancerous cells from cancerous cells based on morphology alone. Additionally, the difference between precancerous cells and cancerous cells in regard to biological behaviour remains unclear. We previously cultured primary normal uterine cervical keratinocytes from human normal cervical tissue and cervical precancerous cells that were naturally infected with human papillomavirus from small-sized neoplastic cervical tissues. Here, we extended our study to further observe the in vitro proliferative characteristics of cervical precancerous cells at the cellular and molecular levels. In this study, we found that the growth rate of precancerous cells was significantly faster than that of normal cervical cells and slower than that of Caski cells. However, the proliferative capacity of such precancerous cells was similar to that of cancerous cells of the cervix at the molecular level. These results suggest that the surrounding environment of the cells may play an important role in the development of cervical cancer, which provides an important basis for the further study of precancerous and cancerous lesions of the cervix.

GHET1 acts as a prognostic indicator and functions as an oncogenic lncRNA in cervical cancer.

Gastric carcinoma proliferation enhancing transcript 1 (GHET1) has been suggested to serve as a promising oncogenic lncRNA in various types of human cancer. However, the role of GHET1 remained unknown in cervical cancer. In our study, we found GHET1 expression was markedly elevated in cervical cancer tissue specimens and cell lines compared with adjacent normal cervical tissue specimens and human normal cervical cell line, respectively. Then, we found high expression of GHET1 is a useful biomarker to discriminate cervical cancer tissues from non-tumorous tissues, and associated with advanced clinical stage, lymph node metastasis, distant metastasis and poor histological grade in cervical cancer patients. The survival analysis showed high GHET1 expression was an independent unfavorable prognostic factor in cervical cancer patients. Knockdown of GHET1 expression markedly inhibits cervical cancer cell proliferation, migration, and invasion. The loss-of-function study indicated knockdown of GHET1 expression markedly inhibits cervical cancer cell proliferation, migration, and invasion. In conclusion, GHET1 acts as an oncogenic lncRNA in cervical cancer.

Proliferation of MicroRNA-365 and E74-like Factor 4 in Cervical Cancer Cells and Its Clinical Significance

Objective To investigate the expressions,roles,and clinical significance of microRNA-365(miR-365)and E74-like factor 4(ELF4)in cervical cancer. Methods The expressions of miR-365 in normal cervical tissues(n=34),cervical intraepithelial neoplasia 1(CIN 1)(n=31),cervical intraepithelial neoplasia2-3(CIN 2-3)(n=37),squamous cell carcinoma of the cervix(SCC)(n=33),and three cervical cancer cell lines(C33A cells,Hela cells,and SiHa cells)were detected by real-time quantitative polymerase chain reaction(qPCR).Bioinformatic prediction and luciferase reporter gene assay were performed to verify whether ELF4 was a direct target of miR-365.Western blot and immunohistochemistry were used to detect ELF4 expression in cervical cancer cells and in different pathological cervix tissues.CCK8 assay was used to detect the effect of overexpression or inhibition of miR-365 on the proliferation of cervical cancer cells at different time points.The relationships among the miR-365 expression,ELF4 expression,and clinicopathological parameters of cervical cancer were analyzed by correlation analysis. Results qPCR results showed that compared with the normal cervical cell HcerEpic,the expressions of miR-365 in CIN1,CIN2-3,and cervical cancer tissues gradually decreased with the increased pathologic grade,and its expressions also decreased in different cervical cancer cell lines.The luciferase reporter gene assay confirmed that ELF4 was the direct target of miR-365.Western blot showed that the expression of ELF4 increased in all three cervical cancer cell lines compared with normal cervical epidermal cell(P=0.013,P=0.002,P=0.004).Immunohistochemistry showed that ELF4 expression was up-regulated in CIN and cervical cancer tissues.CCK8 assay showed that overexpression of miR-365 inhibited cell proliferation,while inhibition of miR-365 promoted the proliferation of three cervical cancer cells(P<0.05).Further analysis confirmed that there was a negative correlation between the expression levels of miR-365 and ELF4 in CIN2-3 and SCC(r=-0.351,P=0.045;r=-0.349,P=0.035).Clinical analysis showed that the expressions of both miR-365 and ELF4 were correlated with tumor size,pathological grade,and clinical stage in SCC(all P < 0.05).<b>Conclusion</b> The decreased expression of miR-365 in human cervical cancer cells relieves its inhibitory effect on ELF4,which promotes the proliferation of cervical cancer cells and the formation of tumor.