Decreased expression of miR-132 in CRC tissues and its inhibitory function on tumor progression

AbstractObjectives Colorectal cancer (CRC) is among the most common types of malignancies in the worldwide, and microRNAs (miRNAs) emerge as key regulators in carcinogenesis and tumor progression. Here we intended to address the expression and function of miR-132 in CRC cells. Methods Paired CRC tissues and several established cell lines were firstly collected. We performed qPCR to detect the expression of miR-132 in these tissues and cell lines. Cell proliferation and apoptosis were respectively monitored by CCK-8 assay and Annexin-V/PI staining followed by flow cytometry, after miR-132 was transiently overexpressed in RKO cells. Afterwards, Luciferase reporter assays were performed to examine the targeting of YAP1 by miR-132. Finally, qPCR and western blotting were also carried out to validate this targeting. Results MiR-132 was significantly decreased in CRC and its overexpression in RKO cells exerted tumor suppressing effects, including cell growth arrest and apoptosis promotion. Additionally, we proved that miR-132 could negatively regulate the expression of YAP1. Conclusion Our findings suggested that miR-132 was downregulated in CRC, and played as a tumor suppressor to inhibit cell proliferation and induce apoptosis. And these anti-tumor activities might be related with the targeting of YAP1 by miR-132.

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LncRNA LINC00858 enhances cervical cancer cell growth through miR-3064-5p/ VMA21 axis

Cancer Biomarkers ◽

10.3233/cbm-200033 ◽

2021 ◽

pp. 1-11

Author(s):

Min Wei ◽

Youguo Chen ◽

Wensheng Du

Keyword(s):

Cervical Cancer ◽

Cell Proliferation ◽

Cell Apoptosis ◽

Luciferase Reporter ◽

Cancer Cell Growth ◽

Protein Coding ◽

Gynecological Malignancy ◽

Promising Therapeutic Target ◽

Proliferation And Apoptosis ◽

Reporter Assays

BACKGROUND: Cervical cancer (CC) is the most common form of gynecological malignancy. Long intergenic non-protein coding RNA 858 (LINC00858) has been identified to participate in multiple cancers. However, the role and mechanism of LINC00858 in CC cells are still elusive. AIM: The aim of this study is to explore the biological functions and mechanisms of LINC00858 in CC cells. METHODS: RT-qPCR analysis was used to examine the expression of LINC00858 in CC cells. EdU and colony formation assay were utilized to assess cell proliferation. TUNEL assay and flow cytometry assay were conducted to assess cell apoptosis. The mechanism regarding LINC00858 was certified through RNA pull down, RIP and luciferase reporter assays. RESULTS: The up-regulated LINC00858 was detected in CC cells. Reduction of LINC00858 effectively subdued CC cells proliferation and stimulated cell apoptosis. LINC00858 was determined to bind with miR-3064-5p and up-regulate VMA21 in CC cells. In rescue assays, miR-3064-5p down-regulation and VMA21 up-regulation were able to counteract the effect caused by LINC00858 decrease on CC cell proliferation and apoptosis. CONCLUSION: LINC00858 enhances cell proliferation, while restraining cell apoptosis in CC through targeting miR-3064-5p/VMA21 axis, implying that LINC00858 may serve as a promising therapeutic target for CC.

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Aberrant expression of hsa_circ_0025036 in lung adenocarcinoma and its potential roles in regulating cell proliferation and apoptosis

Biological Chemistry ◽

10.1515/hsz-2018-0303 ◽

2018 ◽

Vol 399 (12) ◽

pp. 1457-1467 ◽

Cited By ~ 4

Author(s):

Shujun Wu ◽

Hui Li ◽

Chunya Lu ◽

Furui Zhang ◽

Huaqi Wang ◽

...

Keyword(s):

Cell Proliferation ◽

Lung Adenocarcinoma ◽

Luciferase Reporter ◽

Clinicopathological Parameters ◽

Circular Rnas ◽

Aberrant Expression ◽

Close Link ◽

Adenocarcinoma Cells ◽

Proliferation And Apoptosis ◽

Reporter Assays

AbstractAs the most common histological subtype of lung cancer, lung adenocarcinoma remains a tremendous risk to public health, which requires ceaseless efforts to elucidate the potential diagnostic and therapeutic strategies. Circular RNAs (circRNAs) have been identified with emerging roles in tumorigenesis and development. Our preliminary work noticed that hsa_circ_0025036 was significantly upregulated in lung adenocarcinoma tissues. However, its specific roles in lung adenocarcinoma remain unclear. The results in this study revealed that hsa_circ_0025036 existed as a circular form and was aberrantly upregulated in lung adenocarcinoma tissues via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Its expression level exhibited a close link with aggressive clinicopathological parameters including cancer differentiation, TNM stage and lymph node metastasis. hsa_circ_0025036 knockdown significantly suppressed cell proliferation and promoted cell apoptosis in A549 and Calu-3 cells. Moreover, hsa_circ_0025036/miR-198/SHMT1&TGF-αaxis was identified via bioinformatics analysis and Dual-Luciferase Reporter assays. miR-198 inhibitors reversed the function of hsa_circ_0025036 knockdown. hsa_circ_0025036 knockdown exerted similar effects with miR-198 upregulation on cell proliferation and apoptosis. In conclusion, we demonstrate that hsa_circ_0025036 regulates cell proliferation and apoptosis in lung adenocarcinoma cells probably via hsa_circ_0025036/miR-198/SHMT1&TGF-αaxis. hsa_circ_0025036 may serve as a potential prognostic biomarker and a therapeutic target for lung adenocarcinoma.

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miR-1 Inhibits Cell Growth, Migration, and Invasion by Targeting VEGFA in Osteosarcoma Cells

Disease Markers ◽

10.1155/2016/7068986 ◽

2016 ◽

Vol 2016 ◽

pp. 1-8 ◽

Cited By ~ 15

Author(s):

Junjie Niu ◽

Yibao Sun ◽

Qiaoge Guo ◽

Dongju Niu ◽

Bo Liu

Keyword(s):

Cell Proliferation ◽

Cell Lines ◽

U2os Cell ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Small Noncoding Rnas ◽

Normal Tissues ◽

Inhibitory Function ◽

Tumor Tissues ◽

U2os Cells

microRNAs (miRNAs) are small noncoding RNAs and have been shown to play a crucial role in the osteosarcoma (OS) tumorigenesis and progression. VEGFA is a key regulator of angiogenesis and plays an important role in regulation of tumor metastasis. The objective of this study was to determine whether VEGFA was involved in miR-1-mediated suppression of proliferation, migration, and invasion of OS cells. The expression levels of miR-1 were significantly lower in OS tumor tissues than those in adjacent normal tissues and in SAOS-2 and U2OS cell lines compared to a normal osteoblast (NHOst) cell line. VEGFA was upregulated in OS tumor tissues and SAOS-2 and U2OS cell lines. The results of CCK-8 assay and transwell assay showed that miR-1 acted as a tumor suppressor by inhibiting cell proliferation, migration, and invasion in U2OS cells. Dual luciferase reporter assay demonstrated that VEGFA was a direct and functional target gene of miR-1. miR-1 directly inhibits the protein expression of VEGFA via its 3′-UTR. Knockdown of VEGFA by siRNA inhibited proliferation, migration, and invasion of U2OS cells. Our study suggested the potential inhibitory function of miR-1 in OS cell proliferation, migration, and invasion via inhibiting VEGFA.

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MiR-34c regulates the proliferation and apoptosis of lung cancer cells

Clinical & Investigative Medicine ◽

10.25011/cim.v43i4.34997 ◽

2020 ◽

Vol 43 (4) ◽

pp. E56-62

Author(s):

Xianliang Jiang ◽

Ming Li ◽

Li Kei

Keyword(s):

Lung Cancer ◽

Cell Proliferation ◽

Cancer Cells ◽

Cell Lines ◽

Active Role ◽

Lung Cancer Cells ◽

Luciferase Reporter ◽

Normal Tissues ◽

Qrt Pcr ◽

Proliferation And Apoptosis

Purpose: As miR-34c acts as a tumor suppressant for multiple cancers, the purpose of this study was to investigate that role that miR-34c plays in the proliferation and apoptosis of lung cancer. Methods: The expression of miR-34c in 600 patients with lung cancer was quantitatively analyzed with real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) technology and correlated to clinical pathological parameters. The CCK-8 analysis and flow cytometry were carried out to detect cell proliferation and apoptosis in miR-34c-mimic transfected cell lines. Moreover, the regulation of miR-34c to interleukin-6 (IL-6) in cell lines was detected by western blot, qRT-PCR and dual-luciferase reporter assay. Results: The expression of miR-34c was downregulated in lung cancer compared with adjacent normal tissues. The expression level of miR-34c was linked to stromal invasion. Furthermore, overexpressing miR-34c played an active role in effectively inhibiting cell proliferation and inducing apoptosis. In addition, a significant inverse relationship was exhibited between the expression of miR-34c and IL-6 in tumor tissues. Conclusion: At the molecular level, IL-6 can be used as a direct target of miR-34c in the treatment of lung cancer cells and miR-34c can be used as an effective biomarker and therapeutic target for lung cancer.

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Mir-573 regulates cell proliferation and apoptosis by targeting Bax in human degenerative disc cells following hyperbaric oxygen treatment

10.21203/rs.3.rs-88633/v1 ◽

2020 ◽

Author(s):

Song-Shu Lin ◽

Chi-Chien Niu ◽

Li-Jen Yuan ◽

Tsung-Ting Tsai ◽

Po-Liang Lai ◽

...

Keyword(s):

Cell Proliferation ◽

Hyperbaric Oxygen ◽

Caspase 3 ◽

Target Genes ◽

Vital Role ◽

Luciferase Reporter ◽

Nucleus Pulposus Cells ◽

Caspase 9 ◽

Proliferation And Apoptosis ◽

Reporter Assays

Abstract Background: MicroRNA (miRNA) plays a vital role in the intervertebral disc (IVD) degeneration. The expression level of miR-573 was downregulated whereas Bax was upregulated notably in human degenerative nucleus pulposus cells (NPCs). In this study, we aimed to investigate the role of miR-573 in human degenerative NPCs following hyperbaric oxygen (HBO) treatment. Methods: NPCs were separated from human degenerated IVD tissues. The control cells were maintained in 5% CO2/95% air and the hyperoxic cells were exposed to 100% O2 at 2.5 atmospheres absolute. MiRNA expression profiling was performed via microarray and confirmed by real-time PCR, and miRNA target genes were identified using bioinformatics and luciferase reporter assays. The mRNA and protein levels of Bax were measured. The proliferation of NPCs were detected using MTT assay. The protein expression levels of Bax, cleaved caspase 9, cleaved caspase 3, pro-caspase 9 and pro-caspase 3 were examined.Results: Bioinformatics analysis indicated that the 3′ untranslated region (UTR) of the Bax mRNA contained the “seed-matched-sequence” for hsa-miR-573, which was validated via reporter assays. MiR-573 was induced by HBO and simultaneous suppression of Bax was observed in NPCs. Knockdown of miR-573 resulted in upregulation of Bax expression in HBO-treated cells. In addition, overexpression of miR-573 by HBO increased cell proliferation and coupled with inhibition of cell apoptosis. The cleavage of pro‑caspase 9 and pro‑caspase 3 was suppressed while the levels of cleaved caspase 9 and caspase 3 were decreased in HBO-treated cells. Transfection with anti-miR-573 partly suppressed the effects of HBO. Conclusion: Mir-573 regulates cell proliferation and apoptosis by targeting Bax in human degenerative NPCs following HBO treatment.

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Long noncoding RNA SNHG14 regulates ox-LDL-induced atherosclerosis cell proliferation and apoptosis by targeting miR-186-5p/WIPF2 axis

Human & Experimental Toxicology ◽

10.1177/0960327120940363 ◽

2020 ◽

Vol 40 (1) ◽

pp. 47-59

Author(s):

Z Tao ◽

Z Cao ◽

X Wang ◽

D Pan ◽

Q Jia

Keyword(s):

Cell Proliferation ◽

Cell Lines ◽

Noncoding Rna ◽

Low Density Lipoprotein ◽

Density Lipoprotein ◽

Cell Counting ◽

Luciferase Reporter ◽

Interacting Protein ◽

Downstream Target ◽

Proliferation And Apoptosis

To investigate the role of small nucleolus RNA host gene 14 (SNHG14) in the progression of atherosclerosis (AS), bioinformatics analysis, and other relevant experiments (cell counting kit-8, flow cytometry, quantitative real-time polymerase chain reaction, luciferase reporter, RNA immunoprecipitation, RNA pull-down, and western blot assays) were done. The current study revealed that SNHG14 level was high in the serum of AS patients and oxidized low-density lipoprotein (ox-LDL)-induced AS cell lines. Besides, we found that SNHG14 accelerated cell proliferation while inhibited cell apoptosis in ox-LDL-induced AS cell lines. Next, SNHG14 was confirmed to be a sponge for miR-186-5p in AS cells, and it was validated that SNHG14 regulated AS cell proliferation and apoptosis by sponging miR-186-5p. Moreover, we uncovered that WAS-interacting protein family member 2 (WIPF2) was a downstream target of miR-186-5p in AS cells. Finally, it was demonstrated that miR-186-5p modulated AS cell proliferation and apoptosis via targeting WIPF2. To conclude, our research disclosed that SNHG14 affected ox-LDL-induced AS cell proliferation and apoptosis through miR-186-5p/WIPF2 axis, which may provide a theoretical basis for the treatment and diagnosis of AS.

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MiR-566 mediates cell migration and invasion in colon cancer cells by direct targeting of PSKH1

Cancer Cell International ◽

10.1186/s12935-019-1053-1 ◽

2019 ◽

Vol 19 (1) ◽

Cited By ~ 1

Author(s):

Ying Zhang ◽

Siqi Zhang ◽

Jian Yin ◽

Ruisi Xu

Keyword(s):

Cell Proliferation ◽

Cell Growth ◽

Cell Survival ◽

Cell Lines ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Invasion And Migration ◽

Reporter Assays ◽

And Migration ◽

Colon Epithelial Cell

Abstract Background Colorectal cancer (CRC), a common malignancy worldwide, and microRNAs (miRs) have been suggested to play roles in the disease. MiR-566 expression has been shown to be reduced in CRC, but its functions and mechanisms are still unclear. Methods Cell viability was assessed by using the CellTiter 96 AQueous One Solution Cell Proliferation kit. Cell proliferation was measured with MTT assay. Cell metastasis were measured by transwell assay. Luciferase reporter assays was used to confirm the target of MiR-566. PSKH1 expression was measured by RT-PCR and western blot. Results In the present study, we first observed that miR-566 was expressed in several CRC cell lines (SW480, SW620, LoVo, HT29 and Caco-2) at low levels compared to control colon epithelial cell lines (FHC). Further study showed that miR-566 overexpression suppressed cell survival and impeded cell proliferation, whereas inhibition of its expression enhanced cell survival and proliferation. Transwell assays showed that cell invasion and migration were reduced in cells overexpressing miR-566 and increased in those with inhibition of miR-566. Further analysis confirmed that PSKH1 is a target of miR-566. MiR-566 overexpression significantly inhibited PSKH1 expression and reintroduction of PSKH1 partially reversed the effects of miR-566 on CRC cell growth and metastasis in SW480 and Caco-2 cells. Conclusions Taken together, the data show that CRC cell growth and metastasis can be significantly suppressed by miR-566 through targeting PSKH1.

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circRAE1 promotes colorectal cancer cell migration and invasion through modulating miR-338-3p/TYRO3 axis

10.21203/rs.3.rs-19688/v1 ◽

2020 ◽

Author(s):

Jiabin Du ◽

Jianhua Xu ◽

Junxing Chen ◽

Weinan Liu ◽

Pengcheng Wang ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Proliferation ◽

Cell Lines ◽

Cancer Biology ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Circular Rnas ◽

Advanced Tumor Stage ◽

Advanced Tumor ◽

Reporter Assays

Abstract Background: Growing evidences have revealed that long non-coding RNAs (lncRNAs) including circular RNAs (circRNAs) involve in numerous carcinogenesis. However, the roles of circRNAs in the cancer biology of colorectal cancer (CRC) remain vague. Methods: qRT-PCR and western-blot were used to detecte the circRAE1 levels in CRC tissues and CRC cell lines. Cell proliferation, migration and invasion were detected using wound healing assays, and transwell assays. The interaction between circRAE1 and miR-338-3p and TRYO3 was confirmed by dual-luciferase reporter assays. Results: We uncovered that a novel circRNA Hsa_circ_0060967 (also known as circRAE1) was remarkably increased in CRC tissues, and high circRAE1 level was positively associated with advanced tumor stage, lymph node metastasis, and tumor size. Loss-of-function assay indicated that circRAE1 accelerated cell proliferation, migration and invasion. Besides, miR-338-3p , lowly expressed in CRC tissues and CRC cell lines. dual-luciferase reporter assays showed that circRAE1 could sponge miR-338-3p, which targeted TRYO3 in CRC cells. Furthermore, overexpression of circRAE1 could recue the impaired migration and invasion triggered by miR-338-3p mimics or si-TYRO3 in CRC cells and vice versa. Conclusion : We figured out the network of circRAE1, miR-338-3p, and TYRO3 in CRC cells and revealed that increased circRAE1 served as an oncogene through sponging miR-338-3p, resulting in upregulated TYRO3 expression, which suggested that circRAE1 would be a potential therapeutic target and diagnostic marker for CRC treatment.

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MicroRNA-873 Promotes Cell Proliferation, Migration, and Invasion by Directly Targeting TSLC1 in Hepatocellular Carcinoma

Cellular Physiology and Biochemistry ◽

10.1159/000489594 ◽

2018 ◽

Vol 46 (6) ◽

pp. 2261-2270 ◽

Cited By ~ 12

Author(s):

Guoyong Han ◽

Long Zhang ◽

Xuhao Ni ◽

Zhiqiang Chen ◽

Xiongxiong Pan ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Proliferation ◽

Cell Lines ◽

Primary Liver Cancer ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Migration Assay ◽

G1 Phase Arrest ◽

Reporter Assays ◽

Novel Target

Background/Aims: Hepatocellular carcinoma (HCC) is the most common type of primary liver cancer and has the third highest mortality rate among all cancers. MicroRNAs are a class of endogenous, single-stranded short noncoding RNAs. The purpose of this study was to study the role of microRNA-873 in HCC. Methods: The expression of miRNA-873 and tumor suppressor in lung cancer 1 (TSLC1) in HCC tissues and cell lines was detected by real-time quantitative RT-PCR (RT-qPCR) or western blot. A CCK-8 assay was used to examine cell proliferation; flow cytometry was used to assess the cell cycle; the Transwell migration assay was used to test for metastasis. Luciferase assays were performed to assess whether TSLC1 was a novel target of miRNA-873. Results: We showed that miRNA-873 was upregulated in HCC tissues and cell lines compared with the normal control. Knockdown of miRNA-873 inhibited the growth and metastasis of HepG2 and accelerated G1 phase arrest, while overexpression of miRNA-873 had the opposite effect. The dual-luciferase reporter assays revealed that TSLC1 was a novel target of miRNA-873. Further study showed that TSLC1 was decreased in HCC tissues and cell lines. There was a negative correlation between the expression levels of TSLC1 and miRNA-873. The effect of miRNA-873 overexpression was neutralized by TSLC1. We also found that miRNA-873 activated the PI3K/AKT/mTOR signaling pathway and promoted HCC. Conclusions: Our data demonstrated that miRNA-873 promoted HCC progression by targeting TSLC1 and provided a new target for the therapy of HCC.

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circRAE1 promotes colorectal cancer cell migration and invasion through modulating miR-338-3p/TYRO3 axis

10.21203/rs.3.rs-19688/v3 ◽

2020 ◽

Author(s):

Jiabin Du ◽

Jianhua Xu ◽

Junxing Chen ◽

Weinan Liu ◽

Pengcheng Wang ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Proliferation ◽

Cell Lines ◽

Cancer Biology ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Circular Rnas ◽

Advanced Tumor Stage ◽

Advanced Tumor ◽

Reporter Assays

Abstract Background: Growing evidences have revealed that long non-coding RNAs (lncRNAs) including circular RNAs (circRNAs) involve in numerous carcinogenesis. However, the roles of circRNAs in the cancer biology of colorectal cancer (CRC) remain vague. Methods: qRT-PCR and western-blot were used to detecte the circRAE1 levels in CRC tissues and CRC cell lines.,Cell proliferation, migration and invasion were detected using wound healing assays, and transwell assays. The interaction between circRAE1 and miR-338-3p and TRYO3 was confirmed by dual-luciferase reporter assays. Results: We uncovered that a novel circRNA Hsa_circ_0060967 (also known as circRAE1) was remarkably increased in CRC tissues, and high circRAE1 level was positively associated with advanced tumor stage, lymph node metastasis, and tumor size. Loss-of-function assay indicated that circRAE1 accelerated cell proliferation, migration and invasion. Besides, miR-338-3p , lowly expressed in CRC tissues and CRC cell lines. dual-luciferase reporter assays showed that circRAE1 could sponge miR-338-3p, which targeted TRYO3 in CRC cells. Furthermore, overexpression of circRAE1 could recue the impaired migration and invasion triggered by miR-338-3p mimics or si-TYRO3 in CRC cells and vice versa. Conclusion : we figured out the network of circRAE1, miR-338-3p, and TYRO3 in CRC cells and revealed that increased circRAE1 served as an oncogene through sponging miR-338-3p, resulting in upregulated TYRO3 expression, which suggested that circRAE1 would be a potential therapeutic target and diagnostic marker for CRC treatment.

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