Number Of Invasive Cells Research Articles

Long non-coding RNA LINC00511 promotes proliferation, invasion, and migration of non-small cell lung cancer cells by targeting miR-625-5p/GSPT1.

BackgroundLung cancer is a malignant tumor with a high rate of mortality and metastasis. Recently, extensive research has shown that long non-coding RNAs (lncRNAs) play a crucial role in the development and progression of non-small cell lung cancer (NSCLC). In this paper, we aimed to explore the impact of long intergenic non-coding RNA 00511 (LINC00511) on the development and metastasis of NSCLC.MethodsA dataset containing 501 lung squamous cell carcinoma (LUSC) samples and 49 normal samples was downloaded from The Cancer Genome Atlas (TCGA). The differential gene expression and prognostic potential of LINC00511 in LUSC were analyzed by “limma” in R software. Samples of tumor tissues and normal tissues from 67 patients with NSCLC were obtained, along with clinical features. NSCLC cell proliferation, cell cycle, migration, and invasion were detected by LINC00511 knockdown with Cell Counting Kit-8 (CCK-8), flow cytometry, wound-healing assay, and Transwell experiment. The regulatory relationship between LINC00511 and microRNA (miR)-625-5p, or between miR-625-5p and G1 to S phase transition 1 (GSPT1), was detected by luciferase reporter gene assay. LINC00511, miR-625-5p, and GSPT1 expression in tumor and normal tissues and cells was determined by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot. A xenograft experiment in nude mice was performed. Ki67 and GSPT1 expression in the tumor tissues of the nude mice was assessed by immunohistochemistry.ResultsLINC00511 expression was clearly higher in the tumor tissues of the NSCLC patients than in normal tissues (P<0.001). High LINC00511 expression was related to larger tumor size, positive lymph node metastasis, advanced TNM stage, and a lower 5-year survival rate. Compared with those of the shNC group, the NSCLC cells of the shLINC00511 group had a prominently lower optical density (OD) 450 value at 72 h, a lower percentage of cells in S phase, a higher relative wound width, and a lower invasive cell number (P<0.01 or P<0.001). LINC00511 promoted GSPT1 expression via suppressing miR-625-5p. Compared with those of the shNC group, the nude mice of the shLINC00511 group had a much lower subcutaneous tumor volume and weight (P<0.05 or P<0.001).ConclusionslncRNA LINC00511 promotes proliferation, invasion, and migration of NSCLC cells by targeting miR-625-5p/GSPT. LINC00511 may be a potential diagnostic marker and therapeutic target for NSCLC.

AS1 expression in prostate cancer and its effects on proliferation and invasion of prostate cancer cells.

This paper aimed at investigating AS1 expression in prostate cancer (PCa) and its effects on the proliferation and invasion of prostate cancer cells (PCCs). The prostate tissues and the matched adjacent normal prostate tissues excised and preserved during radical prostatectomy in our hospital were collected. The LncRNA NCK1-AS1 expression was detected. PCa patients were followed up for three years to analyze their prognosis. The correlation of LncRNA NCK1-AS1 expression with clinicopathological features was analyzed. Human normal prostate cells and human PCCs were selected, in which LncRNA NCK1-AS1 expression was tested to screen and then transfect the cells. Cell proliferation, invasion and migration were detected. Cell cycles and apoptosis were analyzed. Compared with the adjacent normal tissues, LncRNA NCK1-AS1 was highly expressed in the prostate cancer tissues. Its expression was remarkably different in those with different stages of TNM and with lymphatic metastasis or not. The prognosis of patients with high LncRNA NCK1-AS1 expression was remarkably poorer than that of those with low expression. Compared with the human normal prostate cells, LncRNA NCK1-AS1 expression in the human PCCs remarkably rose, with the greatest difference in 22Rv1 cells. Compared with the Blank group, cell proliferation and the number of plate cloned cells remarkably reduced in the sh-NCK1-AS1 group. Additionally, in this group, the number of invasive and migratory cells remarkably reduced; the expression of invasion-related protein E-cadherin remarkably rose but that of MMP-2 remarkably reduced; cell cycles were arrested and the expression of cycle-related proteins (CDK4, CDK6, cyclin D1) remarkably reduced; the apoptotic rate and the expression of apoptosis-related protein Bax remarkably rose. LncRNA NCK1-AS1 is highly expressed in PCa, so its down-regulation can inhibit PCCs from proliferating and reduce the number of invasive cells.

Effect and mechanism of circular RNA BICD2 on the biological behavior of oral squamous cell carcinoma cells

Objective: To investigate the effects of circular RNA (circRNA) BICD2 (circ-BICD2) on glutamine metabolism, cell proliferation, migration, invasion and apoptosis of oral squamous cell carcinoma (OSCC) cells and to further explore the possible mechanism. Methods: OSCC cells were purchased from Shanghai Institute of Cellular Cells, Chinese Academy of Sciences. Samples of OSCC tissues and corresponding adjacent tissues were collected from 35 OSCC patients who underwent surgical resection at the First Affiliated Hospital of Zhengzhou University from January 2016 to January 2018. Real-time quantitative PCR (RT-qPCR) and western blot were performed to detect the expression levels of circ-BICD2, miR-296-5p and transgelin2 (TAGLN2) in OSCC tissues and cells. Bioinformatics, dual luciferase report experiment and RNA immunoprecipitation (RIP) experiment were used to determine the targeting relationship between circ-BICD2 and miR-296-5p, miR-296-5p and TAGLN2. According to different transfection oligonucleotides or plasmids, CAL27 and SCC9 cells were divided into si-NC group (transfection si-NC), si-circ-BICD2 group (transfection si-circ-BICD2), si-circ-BICD2+anti-miR-NC group (transfected with si-circ-BICD2 and anti-miR-NC), si-circ-BICD2+anti-miR-296-5p group (transfected with si-circ-BICD2 and anti-miR-296-5p), miR-NC group (transfected with miR-NC), miR-296-5p group (transfected with miR-296-5p mimic), miR-296-5p+pcDNA (transfected with miR-296-5p mimic and pcDNA) and miR-296-5p+TAGLN2 group (transfected with miR-296-5p mimic and pcDNA-TAGLN2). Cell counting kit 8(CCK-8), colony formation experiment, flow cytometry, Scratch healing test and Transwell experiment were applied to detect OSCC cell viability, number of colonies, cycle distribution, apoptosis rate, migration rate and invasive cell numbers. Glutamine (gln) consumption, α-ketoglutaric acid (α-KG) production and adenosine triphosphate (ATP) concentration were also detected by the kit. The expression of cyclinD1 and Glutamine hydrolase (GLS1) proteins of CAL27 and SCC9 cells in each of the groups were detected by western blot. Twelve four-week-old clean BALB/c female nude mice were injected with a single-cell suspension of SCC9 cells into the axillary skin to establish a transplanted tumor model. Twelve transplanted tumor model mice were divided into sh-circ-BICD2 group and sh-NC group. Mice were sacrificed by cervical dislocation at 32 days after injection, the tumors were removed and the tumor weight was tested. Results: The expressions of circ-BICD2 (2.54±0.74) and TAGLN2 (1.86±0.15) were increased (P<0.05), while the expression of MiR-296-5p was decreased in OSCC tissues (P<0.05). Cell viability, clone formation numbers, migration rate, invasive cell numbers, S phase cell ratio, glu consumption, α-KG production, ATP concentration, expression of cyclinD1 and GLS1 proteins of OSCC cells were significantly reduced after interference with circ-BICD2 expression. The apoptosis rate, the proportion of cells in G0-G1 phase and the expression of miR-296-5p were significantly increased (P<0.05). Inhibiting the expression of miR-296-5p coulld reverse the effect of interfering with circ-BICD2 on OSCC cell proliferation, migration, invasion, apoptosis and glutamine metabolism (P<0.05). After overexpression of miR-296-5p, cell viability, clone formation red number, migration rate, number of invasive cells, S phase cell ratio, glu consumption, α-KG production, ATP concentration and expressions of cyclinD1, GLS1 and TAGLN2 proteins in OSCC cells were significant decrease. The rate of apoptosis and the proportion of cells in G0-G1 phase were significantly increased (P<0.05). Compared with overexpression of miR-296-5p, cell viability, clone formation red number, migration rate, number of invasive cells, S phase cell ratio, glu consumption, α-KG production, ATP concentration, cyclinD1, GLS1 and TAGLN2 proteins in OSCC cells after overexpression of miR-296-5p and TAGLN2 were significantly increased, and the apoptosis rate and the proportion of cells in the G0-G1 phase were significantly decreased (P<0.05). Compared with the sh-NC group, the tumor weights of mice in the sh-circ-BICD2 group were significantly reduced (P<0.05). Conclusions: Circ-BICD2 was highly expressed in OSCC cells. Interfering with circ-BICD2 could inhibit the proliferation, migration and invasion of OSCC cells, glutamine metabolism and tumor growth and promote cell apoptosis, which might be relate to the regulation of miR-296-5p/TAGLN2 molecular axis.

NEDD4 E3 ubiquitin protein ligase serves an important role in cutaneous melanoma occurrence and development.

The present study aimed to discuss the effects and relative mechanisms of NEDD4 E3 ubiquitin protein ligase (NEDD4) in cutaneous melanoma (CMM) occurrence and development. Clinical cancer and adjacent normal tissues samples were collected to analyze pathological changes and protein expression of NEDD4. Moreover, small interfering (si)RNA was used to knockdown NEDD4 expression in SK-MEL-2 and Malme-3M cells. Cellular proliferation, apoptosis, invasiveness and migration were examined using colony formation, flow cytometric, Transwell and wound-healing assays, respectively. In addition, the relative mRNA and protein expression levels of NEDD4, notch receptor 1 (Notch1) and PTEN were evaluated via reverse transcription-quantitative (RT-q) PCR and western blotting. It was found that NEDD4 mRNA and protein expression were significantly upregulated (both P<0.01). Following NEDD4-knockdown, colony number was significantly decreased, while the apoptotic rate was significantly increased, the invasive cell number was significantly inhibited and the wound-healing capacity was significantly decreased. Following si-NEDD4 transfection, RT-qPCR and western blotting revealed that NEDD4 and Notch1 mRNA and protein expression levels were significantly downregulated, while those of PTEN were significantly upregulated in the SK-MEL-2 and Malme-3M cell lines. Collectively, the current results suggest that NEDD4-knockdown effectively suppressed CMM biological activity by regulating the Notch1/PTEN pathway in vitro.

Honokiol combined with curcumin sensitizes multidrug-resistant human lung adenocarcinoma A549/DDP cells to cisplatin.

The aim of the present study was to discuss the effects and underlying mechanisms of honokiol (HNK) and/or curcumin (CUR) in sensitization of multidrug-resistant human lung adenocarcinoma A549/DDP cells to cisplatin (DDP). An MTS assay was performed to detect the cytotoxicity of HNK, CUR and DDP in A549 and A549/DDP cells and compare their sensitivity. The A549/DDP cells were then divided into 8 groups: Control, HNK, CUR, DDP, HNK + CUR, HNK + DDP, CUR + DDP and HNK + CUR + DDP. Cell proliferation was measured by MTS assay and colony formation assay, cell apoptosis was detected by flow cytometry, cell invasion was evaluated by Transwell assay and cell migration was determined by a wound healing assay. In order to investigate the possible mechanisms, P-glycoprotein (P-gp) protein expression was measured by western blotting and immunofluorescence assays. The mRNA expression levels of AKT, Erk1/2, cyclin-dependent kinase inhibitor 1 (P21), caspase 3, cleaved caspase 3, caspase 9, cleaved caspase 9, poly (ADP-ribose) polymerase (PARP), cleaved PARP, matrix metalloproteinase (MMP)-2 and MMP-9 were examined by reverse transcription-quantitative (RT-q) PCR assay, and the protein expression levels of phosphorylated (p)-AKT, p-Erk1/2, P21, caspase 3, cleaved caspase 3, caspase 9, cleaved caspase 9, PARP, cleaved PARP, MMP-2 and MMP-9 proteins expression by western blot assay. The MTS assay demonstrated that HNK (5 µg/ml), CUR (10 µg/ml) and DDP (5 µg/ml) had no obvious toxicity to A549/DDP cells, and HNK, CUR and DDP were more sensitive in A549 cells compared with A549/DDP cells. The optimal concentrations of HNK (5 µg/ml), CUR (10 µg/ml) and DDP (5 µg/ml) were chosen to carry out the further experiments. Compared with the control group, no significant change was observed in cell proliferation, apoptosis, migration, invasion and related mRNA and protein expression in HNK, CUR, DDP and HNK + CUR groups. The cell proliferation rate in the HNK + DDP and CUR + DDP groups was significantly suppressed with cell apoptosis significantly increased, respectively. The invasion cell number and wound healing rate of HNK + DDP and CUR + DDP groups were significantly depressed compared with the control group, respectively. Immunofluorescence demonstrated that the nuclear volume of P-gp in HNK + DDP and CUR + DDP groups were significantly downregulated compared with the control group, respectively. The RT-qPCR assay demonstrated that the AKT, Erk1/2 and P21 mRNA expression levels were significantly decreased and cleaved caspase 3, cleaved caspase 9 and cleaved PARP were increased in HNK + DDP and CUR + DDP groups compared with the control group. The western blotting results were consistent with the RT-qPCR results. NK + CUR + DDP had improved effects on A549/DDP compared with HNK + DDP or CUR + DDP group, respectively. HNK and/or CUR could improve the sensitivity of DDP to A549/DDP cell by the regulation of P-gp, inducing apoptosis, and inhibiting migration and invasion via AKT/ERK signal pathway in an in vitro study.

Long Non-Coding RNA F11-Antisense 1 (F11-AS1) Suppresses Ovarian Cancer Biological Activity by Regulating Phosphatase and Tensin Homolog Deleted on Chromosome Ten (PTEN)

Aim: To discuss F11-AS1’s effects and mechanisms in ovarian cancer development. Methods: Evaluating F11-AS1 expression by ISH assay and F11-AS1 mRNA level in difference cell lines by RT-qPCR assay. Using MTT, flow cytometry, transwell and wound healing assay to evaluate SKOV3 cell proliferation, cell apoptosis, invasion and migration. And using WB assay to measure PTEN, p-PI3K, AKT, P53 and MMP-9 proteins expressions. Results: F11-AS1 was significantly down-regulation with stage increasing in cancer tissues (P &lt;0.01, respectively). With F11-AS1 transfection, the SKOV3 cell proliferation rate was significantly depressed with cell apoptosis and G1 phase rate significantly increasing (P &lt;0.001, respectively). And then, invasion cell number and wound healing rate of lncRNA group which transfected with F11-AS1 significantly down-regulation (P &lt;0.001). By WB assay, PTEN and P53 proteins expressions significantly up-regulation and p-PI3K, AKT and MMP-9 proteins expressions were significantly down-regulation (P &lt;0.001). Conclusion: F11-AS1 depresses ovarian cancer biological activity by regulating PTEN by vitro study.

Serine/Arginine Repetitive Matrix 2 Antisense RNA 1 Negatively Regulates miR-370-3p and Promotes Hyperplasia, Migration, and Aggression of the Colon Cancer Cell Line

The purpose of this study is to explore the effect and possible machine-processing of the long non-coding RNA (lncRNA) SRRM2-AS1 in the development and pathogenesis of colorectal cancer. LncRNA plays an important role in tumorigenesis and development. LncRNA can regulate gene transcription and translation, cell proliferation, differentiation and apoptosis by affecting gene expression pathways of various coding proteins. SRRM2-AS1 is a kind of lncRNA. Studies have confirmed that the expression of SRRM2-AS1 is increased in colon adenocarcinoma tissues of colon cancer patients and is closely related to the prognosis of patients. However, the influence and molecular mechanism of SRRM2-AS1 on the malignant biological behavior of colon cancer cells are no yet clear. SRRM2-AS1 may interact with miR-370-3p. Studies have confirmed that overexpression of miR-370-3p can inhibit the proliferation and epithelial-mesenchymal transition of colon cancer cells in vitro. However, it is not yet clear whether SRRM2-AS1 can target miR-370-3p to affect the occurrence and development of tumors. In this study, RT-qPCR was employed to detect levels of SRRM2-AS1 and miRNA-370-3p in carcinoma tissues and corresponding paracarcinoma tissues from 41 patients with colon cancer. SW1116 colon cancer cells were cultured in vitro and separated into 4 groups: (1) si-NC group, (2) si-SRRM2-AS1 group, (3) si-SRRM2-AS1+anti-miRNA-NC group, and (4) si-SRRM2-AS1+anti-miRNA-370-3p group. The CCK-8 assay and colony formation experiment was employed to gauge cell proliferation. The scratch test was used to detect cell migration while the transwell assay was used to detect cell invasion. Finally, Western blot analysis was employed to detect levels of Ki67, E-cadherin, and N-cadherin proteins in colorectal cancer cells. The dual-luciferase reporter gene experiment verified that SRRM2-AS1 regulates miRNA-370-3p. The study found that compared to paracarcinoma tissue, levels of SRRM2-AS1 in colon cancer tissues was increased (P &lt; 0.05). Compared to the si-NC group, the SW1116 cell OD value, number of colonies formed, scratch healing rate, number of invasive cells, and expression levels of Ki67 and N-cadherin protein in the si-SRRM2-AS1 group were all decreased (P &lt; 0.05). However, E-cadherin protein levels were elevated (P &lt; 0.05). SRRM2-AS1 negatively regulates levels of miRNA-370-3p in SW1116 cells. Compared to the si-SRRM2-AS1+anti-miRNA-NC group, SW1116 cell OD value, number of colonies formed, scratch healing rate, number of invasive cells, and Ki67 and N-cadherin protein levels were increased (P &lt; 0.05) in the si-SRRM2-AS1+anti-miRNA-370-3p group. Conversely, E-cadherin protein levels were decreased (P &lt; 0.05). These findings indicate that SRRM2-AS1 is predominately expressed in cancerous colon tissues. Attenuating expression of SRRM2-AS1 may curb the hyperplasia of colon carcinoma cell line SW1116 and promote cell apoptosis by regulating miRNA-370-3p expression.

Effects of ascites derived exosomes on the stemness and invasion ability of ovarian cancer stem-like cell

Objective: To investigate the effects of ovarian cancer ascites-derived exosomes on the stem cell properties and invasion ability of ovarian cancer stem-like cell (OCS-LC). Methods: (1) A2780 cells were induced into OCS-LC in serum-free medium, and authenticating their stem-like properties by sphere-forming test, differentiation test and CD133 marker detection. (2) Exosomes from ascites and A2780 cell were extracted by ultracentrifugation, then authenticating them. The morphology of exosomes was observed by the transmission electron microscope, exosome particle size was measured by nanoparticle tracking analysis (NTA). The expressions of heat shock protein 70 (HSP-70), CD63 and CD9 were detected by western blot. (3) The exosomes from ovarian cancer ascites and tumor cell supernate were co-cultured with OCS-LC. The groups were divided into control group, ascites-derived exosomes (ADE) group (ADE+OCS-LC group), and cells-derived exosomes (CDE) group (CDE+OCS-LC group). The sphere-forming ability was evaluated by sphere-forming cycle, maximum sphere diameter and sphere-forming rate of each group; the expression of CD133 was detected by immunofluorescence staining under microscope; quantitative real-time (qRT)-PCR was used to detected the expression levels of octamer-4 (Oct-4), Nanog mRNA of the signature genes in the stem cells of each group; the metastasis ability of each group was measured by transwell assay. Results: (1) Identification of OCS-LC: sphere-forming experiment showed that the suspension of OCSC single cells was grown in serum-free medium in secondary sphere-forming. Differentiation function experiment showed that OCS-LC were differentiated into adherent A2780 cells by changing the growth mode in serum containing medium. Flow cytometry showed that the proportion of CD133 positive (CD133+) cells in OCS-LC group was (18.9±0.9)%, significantly higher than that of control group (0.6±0.5)% (t=38.570, P<0.01). (2) Under transmission electron microscope, clear lipid bilayer structure was observed in ADE and CDE, and one side presented a concave hemispheric or cup like structure. NTA showed that the diameter of exosomes mainly ranged from 30 to 100 nm, with an average diameter of 67.2 nm. Western blot analysis showed that the specific protein molecules HSP-70, CD63 and CD9 were positive. (3) Three groups' OCS-LC could continue to grow into spheres, and the group of ADE+OCS-LC showed two growth modes, most of the cells continued to grow into spheres, while a small part of cells grew in adherent differentiation. The sphere-forming rate of OCS-LC in the control group, ADE+OCS-LC group, and CDE+OCS-LC group were (1.05±0.20)%, (4.15±0.10)%, and (10.45±0.25)%, the sphere-forming cycle of OCS-LC in the three groups were (15.3±1.5), (10.3±0.6), and (6.7±0.6) days, and the maximum diameters of OCS-LC were (100.3±3.2), (145.2±5.1) and (170.0±2.1) μm, respectively. And the differences were statistically significant (all P<0.05). After co-culture of exosomes with OCS-LC, the sphere-forming ability of cells in the group of CDE+OCS-LC was prior to ADE+OCS-LC group (all P<0.05). Immunofluorescence staining showed that the number of CD133 green fluorescent chromophore cells in OCS-LC groups [(46.2±2.1)%, (58.4±2.2)%] was significantly higher than that in the control group [(26.6±1.5)%] after the addition of exosomes in co-culture, the positive rate of CD133 was higher than that in the control group(F=187.588, P<0.05). The qRT-PCR results showed that the expression levels of Oct-4 mRNA in ADE+OCS-LC and CDE+OCS-LC groups were 3.46±0.24, 4.03±0.31, compared with that in control group (1.04±0.12), the differences were statistically significant (F=134.932, P<0.05). The mRNA expression levels of Nanog were 1.57±0.32, 2.66±0.15, which were significantly higher than that in the control group (1.00±0.07), and the differences were statistically significant (F=49.329, P<0.05). And the expression of both in CDE+OCS-LC group increased more significantly than ADE+OCS-LC group (all P<0.05). The number of invasive cells in the three groups of OCS-LC were: control group 30±5, ADE+OCS-LC group 102±4, CDE+OCS-LC group 210±7, and there were statistically significant differences among three groups (F=820.800, P<0.05). Compared with the control group, the number of invaded cells in the co-culture group were significantly increased (P<0.05), and the CDE+OCS-LC group had the higher cell invasion ability then the ADE+OCS-LC group (t=23.202, P<0.05). Conclusions: Exosomes derived from ovarian cancer ascites could enhance and maintain the stemness of OSC-LC, and promote the invasion of tumor cells. Moreover, CDE is superior to ADE.

HER2 in situ hybridization test in breast cancer: quantifying margins of error and genetic heterogeneity.

The aim of the present study was to evaluate the effect of counting increasing number of invasive cancer cells in the result of the HER2 in situ hybridization (ISH) test in breast cancer as well as to compare two different approaches of measuring genomic heterogeneity (single cell and population based). A cohort of 100 consecutive breast cancer cases (primary and metastatic) were evaluated for HER2 gene amplification with bright-field ISH. The evaluation of the samples included scoring 20 nuclei, in five different areas, measuring the margins of error for each case. Genomic heterogeneity (GH) was defined by the 2018 ASCO/CAP guideline as a discrete population of tumor cells with HER2 amplification. We also evaluated GH as single tumor cells with HER2 amplification. The stabilization of the coefficient of variation of HER2/CEP17 ratio requires about 60 invasive cancer cells. The average margin of error of HER2/CEP17 ratio and of HER2 copy number was 0.40 and 0.53, respectively, when counting 20 cells, decreasing to 0.20 and 0.26 when counting 100 cells. Population GH was observed in 1% of the cases, while single cell GH was observed in 27% of the cases, reaching its maximum value in cases near the thresholds of positivity. Therefore, margins of error in HER2 ISH test are high, and the minimal cell number recommended in current guidelines should be raised to at least 60 cells. Population GH is a rare event and single cell GH is maximal in cases near the thresholds.

Quercetin inhibits invasion and angiogenesis of esophageal cancer cells

BackgroundEsophageal carcinoma has poor prognosis and novel therapies for esophageal carcinoma are urgently needed. Quercetin is a natural flavonoid compound that can be found in many foods. In this study, we investigated the effects of quercetin on invasion and angiogenesis of esophageal cancer cells. MethodsHuman esophageal cancer cell line Eca109 was treated with 5 μg/mL or 10 μg/mL of quercetin. Colony formation assay was performed. Cell migration and invasion were evaluated by wound healing and transwell assays, respectively. Human umbilical vein/vascular endothelium cells (CLR-1730) were treated with Eca109 conditioned medium, and the effects of quercetin on CLR-1730 were evaluated by wound healing and tube formation assays. Protein levels of VEGF-A, MMP9, and MMP2 were determined by Western blotting. ResultsThe ability of colony forming in Eca109 was reduced with the administration of 10 μg/mL quercetin, but there was no difference between the 5 μg/mL quercetin group and control. The migration distance and the number of invasive cells were significantly reduced in the 10 μg/mL quercetin group. At the lower level of quercetin at 5 μg/mL, only the invasion of cells was significantly inhibited. In endothelial cells treated with Eca109 conditioned medium, cell migration and tube forming ability were suppressed. The decreased protein levels of VEGF-A, MMP9, and MMP2 were observed at the 10 μg/mL quercetin group. ConclusionQuercetin suppressed the invasion and angiogenesis of esophageal cancer cells, and the effects were associated with the decreased expression of VEGF-A, MMP2, and MMP9.

Expression of cyclophilin A in oral squamous cell carcinoma and its effect on cell proliferation and invasion

To investigate the expression of cyclophilin A (CyPA) in oral squamous cell carcinoma (OSCC) and explore the effect of downregulating the expression of CyPA gene on the proliferation and invasion of SCC-25 cells. A total of 77 cases of patients with OSCC were selected. The expression levels of CyPA proteins in OSCC and adjacent normal tissues were evaluated. SCC-25 cells were cultured and divided into the CyPA interference sequence group, negative control group, and blank group. The expression levels of CyPA mRNA and protein in cells were detected by using real-time fluorescent quantitative polymerase chain reaction and Western blot, respectively. Cell proliferation was detected by using methyl thiazolyl tetrazolium and plate colony formation assays. Cell invasion was detected by using Transwell assay. The positive expression rate of CyPA protein in OSCC tissues was 76.62%, which was higher than that in adjacent tissues (P<0.05). The positive expression rate of CyPA protein in TNM stage T3+T4, clinical stage Ⅲ+Ⅳ, moderately or poorly differentiated lymph node metastasis was increased (P<0.05). Compared with the negative control and blank groups, the CyPA interference sequence group had decreased relative expression levels of CyPA mRNA and protein (P<0.05); optical density va-lues of cells at 24, 48, 72, and 96 h (P<0.05); and number of cell colonies and invasive cells (P<0.05). The CyPA protein is highly expressed in OSCC tissues, and the downregulation of CyPA gene expression in SCC-25 cells can reduce cell proliferation and inhibit cell invasion.

LINC01006 regulates the proliferation, migration and invasion of hepatocellular carcinoma cells through regulating miR-433-3p/CBX3 axis

Introduction and objectivesLINC01006 has been verified to be correlated with several cancer types, whereas its biological function in hepatocellular carcinoma (HCC) is still elusive. This study aimed to elucidate the specific regulatory mechanism of LINC01006 in the tumorigenesis of HCC. Materials and methodsThe expression of LINC01006, miR-433-3p and CBX3 in HCC tissues and cells was assessed by qRT-PCR or Western blot. MTT, wound-healing, and transwell assays were used to evaluate the effects of LINC01006 on cell viability, migration, and invasion in vitro. A mouse xenograft model was established for in vivo assays. The relations among LINC01006, miR-433-3p, and CBX3 were analyzed by MS2-RNA immunoprecipitation (RIP) and Dual-luciferase reporter (DLR) assays. ResultsThe expression of LINC01006 was up-regulated in HCC tissues and cells. LINC01006 knockdown inhibited the viability, wound healing rate, and invasive cell number of HeP3B and SK-HeP-1 cells, and decreased the tumor volume and weight in a mouse xenograft model. MiR-433-3p was a target of LINC01006, and LINC01006 overexpression inhibited the viability, wound healing rate, and invasive cell number of HeP3B and SK-HeP-1 cells. In addition, CBX3 was a target of miR-433-3p, which was negatively regulated by miR-433-3p. CBX3 overexpression and miR-433-3p inhibition reversed the inhibiting effects of LINC01006 knockdown on the viability, migration, and invasion of HeP3B cells. ConclusionsSilencing of LINC01006 inhibited the viability, migration, and invasion of HCC cells through regulating miR-433-3p/CBX3 axis.

Toosendanin Affects the Occurrence and Development of Prostate Carcinoma Cells DU145 by Regulating Forkhead Box C2-Antisense RNA 1

Prostate carcinoma is a common malignant tumor of the male genitourinary system. Toosendanin can inhibit the biological behavior of a variety of malignant tumor cells (such as ovarian carcinoma, lung carcinoma, and breast carcinoma, etc.), but its effect on the malignant behavior of prostate carcinoma cells and its mechanism are not yet understood. Therefore, this article discusses the influence of toosendanin on the multiplication, apoptosis, migration, and invasion of prostate carcinoma cells and its possible mechanism. Different doses (0.125, 0.25, 0.5 ^M) of toosendanin can reduce the cell viability, number of colonies, number of migrating cells, number of invasive cells, and Bcl-2 protein and FOXC2-AS1 levels of prostate carcinoma cells, as well as increase the apoptosis rate and Bax protein level. Overexpression of FOXC2-AS1 can increase the cell viability, number of colonies formed, number of migrating cells, number of invasive cells, and Bcl-2 protein expression, as well as reduce the rate of apoptosis and Bax protein level after toosendanin treatment of prostate carcinoma cells. It was demonstrated that toosendanin may inhibit the multiplication, migration, and invasion of prostate carcinoma cells and promote its apoptosis by down-regulating FOXC2-AS1 expression.

Dihydromyricetin Inhibits Proliferation, Migration, and Invasion of Pancreatic Cancer Cells by Regulating miR-509-3p/PRMT5 Pathway

To explore the role of dihydromyricetin (DHM) in regulating proliferation, metastasis and infiltration of pancreatic cancer cells, we treated the pancreatic adenocarcinoma cell line SW1990 with 10 µmol/L, 20 µmol/L, and 40 µmol/L DHM. Proliferation was determined by the MTT assay. Cell metastasis and infiltration were determined by the Transwell migration assay. The protein expression levels of cyclin D1, p21, MMP-2, MMP-9, and PRMT5 were determined by Western blot. Quantitative PCR was used to determine expression of miR-509-3p and PRMT5 mRNA. The relationship between miR-509-3p and PRMT5 was analyzed by bioinformatic prediction and the dual luciferase reporter assay. SW1990 cells were transfected with miR-509-3p, si-PRMT5 and anti-miR-509-3p (following treatment with 40 µmol/L DHM) to determine their effects on biological behaviors of cells. The inhibition rate of SW1990 cells, p21 protein and miR-509-3p expression were increased by DHM. Moreover, the number of infiltrating and metastatic cells, protein levels of cyclin D1, MMP-2, MMP-9, and PRMT5, and mRNA levels of PRMT5 were decreased by DHM. miR-509-3p regulates expression of its target PRMT5 mRNA. Inhibition rate of SW1990 cell proliferation and protein level p21 were significantly increased by overexpression of miR-509-3p or knockdown of PRMT5 expression, while the number of invasive cells and protein expression levels of cyclin D1, MMP-2 and MMP-9 were remarkably reduced by miR-509-3p overexpression or PRMT5 knockdown. Inhibition of miR-509-3p counteract the inhibitory effects of DHM on SW1990 cell proliferation and metastasis. Therefore, DHM inhibits proliferation and metastasis of pancreatic carcinoma cells by regulating the miR-509-3p/PRMT5 pathway.

Inhibition of BAMBI reduces the viability and motility of colon cancer via activating TGF-β/Smad pathway in vitro and in vivo.

[This retracts the article DOI: 10.3892/ol.2017.6811.].

Expression of lncRNA MSC-AS1 in hepatocellular carcinoma cell lines and its effect on proliferation, apoptosis, and migration.

This study aimed to explore the expression of long non-coding RNA MSC-AS1 in hepatocellular carcinoma (HCC) cells and its effect on the proliferation, migration, and apoptosis of HCC cells. The expression of MSC-AS1 in HCC cell lines BEL7402, SMMC7721, Huh7, HepG2, MHCC97-H, and normal hepatocyte line L02 was detected by reverse transcriptase polymerase chain reaction. The HCC cells were divided into blank, negative control (NC)-small interfering RNA (siRNA) (transfected with negative siRNA), and MSC-AS1 siRNA (transfected with MSC-AS1 siRNA) groups. Cell counting kit-8 and colony formation assays were used to determine the proliferation, and cell apoptosis, migration, and invasion were detected by flow cytometry, wound healing, and transwell assays, respectively. Western blot was used to detect the expression of related proteins. The expression of MSC-AS1 in HCC cell lines was significantly higher than that in L02. In the MSC-AS1 siRNA group, the proliferation and colony formation of HCC cells were inhibited, whereas the apoptosis rate was significantly higher than that in the blank and NC-siRNA groups. The rate of wound healing and the number of invasion cells in the MSC-AS1 siRNA group were significantly lower than that in the blank and NC-siRNA groups. MSC-AS1 was upregulated in HCC cells, and the downregulation of MSC-AS1 could inhibit cell proliferation, migration, and invasion and promote apoptosis of HCC cells.

Study on Effect of Vitexicarpin on the Biological Behavior of Colorectal Cancer Cells by Circ_0000419/MicroRNA-224

To investigate the effect and molecular mechanism of vitexicarpin on the biological behavior of colorectal cancer cells is the objective of the study. The human epithelial cell line Caco-2 cells were divided into control group, low, middle and high dose groups of vitexicarpin, plasmid cloning DNA-circ_0000419 group, plasmid cloning DNA group, anti-microRNA-224 group, anti-microRNA-224-negative control group, vitexicarpin+small interfering-circ_0000419 group and vitexicarpin+microRNA-224 group. Cell activity was detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide assay. Colony formation number was detected by plate clone formation experiment. Apoptosis was detected by flow cytometry. Scratch test was used to detect the migration distance of cells. Transwell method was used to detect the number of invasive cells; protein expression was detected by western blot; the expression levels of circ_0000419 and microRNA-224 were detected by real-time fluorescence quantitative polymerase chain reaction; dual-luciferase reporter assay was used to detect the targeting relationship between circ_0000419 and microRNA-224. After treatment with low, middle and high doses of vitexicarpin, the viability of Caco- 2 cells decreased, the number of colony formation decreased, the apoptosis rate increased, the migration distance decreased, the number of invasive cells decreased, the expression level of E-cadherin increased, the expression level of N-cadherin decreased, the expression level of circ_0000419 increased and the expression level of microRNA-224 decreased (p<0.05). Overexpression of circ_0000419 or inhibition of miR-224 expression decreased Caco-2 cell viability, decreased colony formation, increased apoptosis rate, decreased cell migration distance, decreased invasive cell number, increased expression of E-cadherin and decreased expression of N-cadherin (p<0.05). Interference with circ_0000419 or overexpression of microRNA-224 reverses the effects of vitexicarpin on proliferation, apoptosis, migration and invasion of Caco-2 cells; circ_0000419 targets the regulation of microRNA-224 expression. Vitexicarpin can inhibit the malignant biological behavior of colorectal cancer cells by circ_0000419/microRNA-224.

Mechanism of Regulating the Proliferation, Migration and Invasion of Cervical Cancer Hela Cells by Circular RNA Circ_0025033 Targeting microRNA 1256

To explore the effect of circular RNA_0025033 on the proliferation, migration and invasion of cervical cancer HeLa cells and its molecular mechanism. The cancer tissues and adjacent tissues of 39 patients with cervical cancer in our hospital from January 2017 to December 2020 were selected. The expression levels of circular RNA_0025033 and microRNA-1256 were detected by Quantitative reverse transcription polymerase chain reaction. Cell counting kit 8 was used to detect cell viability. Plate cloning test was conducted to detect the number of cell clones. Scratch test was carried out to detect cell scratch healing rate. Transwell chamber was used to detect cell invasion. Western blot was adopted to detect protein expression. Dual luciferase reporter assay was carried out to detect the targeting relationship between circular RNA_0025033 and microRNA-1256. The expression level of circular RNA_0025033 in cervical cancer tissues was increased and the expression level of microRNA-1256 was decreased (p<0.05). After inhibiting the expression of circular RNA_0025033 or overexpression of microRNA-1256, cell viability was decreased and the number of cell clone formation was decreased. The healing rate of cell scratches was decreased and the number of invasive cells was decrease. E-cadherin expression was increased and N-cadherin expression was decreased (p<0.05). Circular RNA_0025033 targeted regulation of the expression of microRNA-1256 was carried out; interference with microRNA-1256 expression reversed the inhibition of circular RNA_0025033 on the proliferation, migration and invasion of cervical cancer HeLa cells. Inhibition of circular RNA_0025033 expression may inhibit the proliferation, migration and invasion of cervical cancer HeLa cells through targeted regulation of microRNA-1256.

MicroRNA-377-3p targeting MMP-16 inhibits ovarian cancer cell growth, invasion, and interstitial transition.

BackgroundTo evaluate role of microRNA (miRNA)-377-3p on the remission of ovarian cancer (OC) cell proliferation, invasion, and interstitial transition in vivo and vitro.MethodsSKOV3 cells were used as the object of in vitro research and four-week-old immunodeficient BABL/c female nude mice were used to form the xenograft model. Cell models were constructed by transfecting NC mimics, miR-377 mimic, plasmid cloning DNA (pcDNA), pc-matrix metalloproteinase (MMP)-16, or co-transfecting miR-377 mimic and pc-MMP-16. TargetScan software was used to predict the targeting relationship between miRNA-377-3p and MMP-16 in OC cells. The combination of miRNA-377-3p and MMP-16 was detected by dual luciferase report experiment. miRNA expression levels of miRNA-377-3p and MMP-16 in each transfection group cells were detected by reverse transcription-polymerase chain reaction (RT-PCR). The proliferation of SKOV3 cells were assessed by 5-ethynyl-2'-deoxyuridine (EdU) staining and microtubule formation, while the invasion ability of SKOV3 cells was detected by Transwell assay. Protein expression levels of MMP-16, survivin, Ki67, vascular endothelial growth factor (VEGF), E-cadherin, and N-cadherin were detected by Western blot (WB), and the positive cells of Ki67 and VEGF were detected by immunohistochemistry (IHC).ResultsMMP-16 overexpression markedly increased the EDU-positive cell percentage, upregulated survivin and Ki67 levels, increased the number of invasive cells per field, and enhanced VEGF and N-cadherin expression. Importantly, co-transfection of miRNA-377-3p and MMP-16 reversed these abnormal phenomena. Xenotransplantation mouse models were formed by injecting SKOV-3 cells subcutaneously. Tumor size, tumor volume, and tumor weight were all reduced in the miR-377-3p mimic–transfected group. The results of IHC indicated that Ki67 and VEGF expression were decreased in the miR-377-3p mimic–transfected group.ConclusionsThese findings indicate that miR-377-3p could be a promising therapeutic agent for OC cell growth, invasion, and interstitial transition with MMP-16 being its likely target.

Paclitaxel inhibits proliferation and invasion and promotes apoptosis of breast cancer cells by blocking activation of the PI3K/AKT signaling pathway.

Breast cancer has the highest incidence and mortality among all cancers in women. Paclitaxel (PTX) has a notable therapeutic effect on cancer in clinical practice. To explore the effect and mechanism of PTX on the proliferation, apoptosis and invasiveness of breast cancer cells. MCF-7 cells were treated with PTX (0 μM, 0.01 μM, 0.1 μM, 1 μM) for 48 h. Cell viability was detected using MTT assay and lactate dehydrogenase (LDH) assay; the cell proliferation rate was detected using 5-ethynyl-2'-deoxyuridine (EdU) assay to screen the most effective concentration of PTX. MCF-7 cells were then divided into 5 groups: control group, PTX group, oe-PI3K group, NC-PI3K group, and oe-PI3K+PTX group. Cell apoptosis and cell cycles were detected with flow cytometry; cell invasion was determined using a transwell assay; western blot and quantitative reverse-transcription polymerase chain reaction (qRT-PCR) were used to measure the mRNA and protein expression level of cleaved caspase-3, Bax, Bcl-2, matrix metalloproteinase 9 (MMP-9), vascular endothelial growth factor (VEGF), p-AKT (Thr308), and p-AKT (Ser473). Paclitaxel inhibited cell viability and proliferation in a dose-dependent manner. In the PTX group, the apoptosis rate, the number of cells arrested in the G2/M phase and the expression levels of Cleaved caspase-3 and Bax were increased, but the number of invasive cells and the expression levels of Bcl-2, MMP-9, vascular endothelial growth factor (VEGF), p-AKT (Thr308), and p-AKT (Ser473) were decreased. However, PI3K upregulation can reverse the effects of PTX. Paclitaxel could inhibit MCF-7 cell proliferation and invasion, and promote MCF-7 cell apoptosis by downregulating the expression of p-AKT (Thr308) and p-AKT (Ser473) in the PI3K/AKT signaling pathway.