scholarly journals MiR-199a-5p-HIF-1α-STAT3 positive feed-back loop contributes to the progression of NSCLC

2020 ◽  
Author(s):  
Xingping Yang ◽  
Yuzhen Zheng ◽  
Jian Tan ◽  
Xiansheng Hu ◽  
Renjiang Tian ◽  
...  
Keyword(s):  
A549 Cells ◽  
Bioinformatic Analysis ◽  
Luciferase Assay ◽  
Migration And Invasion ◽  
Flow Cytometry Analysis ◽  
Transwell Assay ◽  
Xenograft Models ◽  
Direct Target ◽  
Induced Apoptosis ◽  
Common Malignancy

Abstract Background Non-small cell lung cancer (NSCLC), is the most common malignancy worldwide. MiR-199a-5p has been reported to play important roles in multiple tumors, inclusive of NSCLC. Whereas, little is definitively known pertaining to its explicit mechanism of action in NSCLC. Methods Expression of miR-199a-5p and HIF-1α mRNA were quantified employing qRT-PCR. H1299 and A549 cells were transiently transfected with miR-199a-5p mimics or inhibitors. Then CCK-8 assays, flow cytometry analysis, Transwell assay were implemented for detecting cell proliferation, cell cycle, apoptosis, migration and invasion of NSCLC cells, respectively. HIF-1α, STAT3 and p-STAT3 expression were detected via Western blotting. Bioinformatic analysis and dual-luciferase assay were performed for monitoring interaction between miR-199a-5p, HIF-1α and STAT3. Xenograft models were established with nude mice for further analyzing Bevacizumab resistance of NSCLC. Results MiR-199a-5p expression was markedly attenuated in NSCLC tissues and cell lines. Overexpression of miR-199a-5p constrained proliferation, migration, invasion but induced apoptosis of NSCLC cells. HIF-1α was identified as a direct target of miR-199a-5p. Further study confirmed a positive feed-back loop between miR-199a-5p, HIF-1α and STAT3. Co-transfection of HIF-1α or STAT3 overexpression plasmids counteracted effects of miR-199a-5p. Further study substantiated that feed-back loop might be in association with the Bevacizumab resistance of NSCLC cells . Conclusion MiR-199a-5p constrained the progression and Bevacizumab resistance of NSCLC by suppressing HIF-1α and STAT3, while HIF-1α/STAT3 axis suppressed the expression of miR-199a-5p, which forms a positive feed-back loop to promote the sustaining progression of NSCLC.

scholarly journals Downregulation of circRNA_100876 Inhibited Progression of NSCLC In Vitro via Targeting miR-636

2020 ◽  
Vol 19 ◽  
pp. 153303382095181
Author(s):  
Jianxiang Song ◽  
Woda Shi ◽  
Zhengya Gao ◽  
Xingchen Liu ◽  
Wencai Wang
Keyword(s):  
Small Cell Lung Carcinoma ◽  
Luciferase Assay ◽  
Migration And Invasion ◽  
Transwell Assay ◽  
Cell Lung Carcinoma ◽  
Downstream Target ◽  
Protein Expressions ◽  
Direct Target ◽  
Related Proteins

Background: Non-small cell lung carcinoma (NSCLC) is a common malignant tumor with poor prognosis. CircRNA-100876 has been considered to be involved in NSCLC. However, the mechanism by which circRNA_100876 mediated the progression of NSCLC remains unclear. Methods: CCK8 assay and immunofluorescence were used to detect cell proliferation. Flow cytometry and transwell assay were performed to analyze cell apoptosis, migration and invasion, respectively. Verification of possible target for circRNA_100876 and related miR-636 were done using luciferase assay. In addition, western blot was performed to detect the protein expressions in NSCLC cells. Results: Silencing of circRNA_100876 notably inhibited the proliferation of NSCLC cells. Moreover, downregulation of circRNA_100876 significantly induce the apoptosis of NSCLC cells via mediation of apoptosis-related proteins. In addition, silencing of circRNA_100876 significantly inhibited migration and invasion of NSCLC cells. MiR-636 was the downstream target of circRNA_100876. Meanwhile, RET was the direct target of miR-636. Finally, circRNA_100876 shRNA2 notably suppressed the progression of NSCLC through PI3K/Akt signaling. Conclusion: CircRNA_100876 knockdown notably suppressed the progression of NSCLC through regulation of miR-636/RET axis, which may serve as a potential target for treatment of NSCLC.

scholarly journals HBP1-mediated transcriptional repression of AFP inhibits hepatoma progression

2021 ◽  
Vol 40 (1) ◽  
Author(s):  
Zhengyi Cao ◽  
Yuning Cheng ◽  
Jiyin Wang ◽  
Yujuan Liu ◽  
Ruixiang Yang ◽  
...  
Keyword(s):  
Transcriptional Regulation ◽  
Transcriptional Repression ◽  
Chinese Herb ◽  
Gene Promoter ◽  
Hepatoma Cells ◽  
Luciferase Assay ◽  
B Virus ◽  
And Migration ◽  
Induced Apoptosis ◽  
Common Malignancy

Abstract Background Hepatoma is a common malignancy of the liver. The abnormal high expression of alpha-fetoprotein (AFP) is intimately associated with hepatoma progress, but the mechanism of transcriptional regulation and singularly activation of AFP gene in hepatoma is not clear. Methods The expression of transcription factor HBP1 and AFP and clinical significance were further analyzed in hepatoma tissues from the patients who received surgery or TACE and then monitored for relapse for up 10 years. HBP1-mediated transcriptional regulation of AFP was analyzed by Western blotting, Luciferase assay, Realtime-PCR, ChIP and EMSA. After verified the axis of HBP-AFP, its impact on hepatoma was measured by MTT, Transwell and FACS in hepatoma cells and by tumorigenesis in HBP1−/− mice. Results The relative expressions of HBP1 and AFP correlated with survival and prognosis in hepatoma patients. HBP1 repressed the expression of AFP gene by directly binding to the AFP gene promoter. Hepatitis B Virus (HBV)-encoded protein HBx promoted malignancy in hepatoma cells through binding to HBP1 directly. Icaritin, an active ingredient of Chinese herb epimedium, inhibited malignancy in hepatoma cells through enhancing HBP1 transrepression of AFP. The repression of AFP by HBP1 attenuated AFP effect on PTEN, MMP9 and caspase-3, thus inhibited proliferation and migration, and induced apoptosis in hepatoma cells. The deregulation of AFP by HBP1 contributed to hepatoma progression in mice. Conclusions Our data clarify the mechanism of HBP1 in inhibiting the expression of AFP and its suppression in malignancy of hepatoma cells, providing a more comprehensive theoretical basis and potential solutions for the diagnosis and treatment of hepatoma.

scholarly journals Alkannin Inhibits the Development of Ovarian Cancer by Affecting miR-4461

2021 ◽  
Vol 2021 ◽  
pp. 1-8
Author(s):  
Yaowen Wang ◽  
Jingfang Zhang ◽  
Feipeng Wang ◽  
Wenping Chen ◽  
Jie Ma ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Flow Cytometry ◽  
Cell Viability ◽  
Migration And Invasion ◽  
Flow Cytometry Analysis ◽  
Antibacterial Effects ◽  
Transwell Assay ◽  
Ovarian Cells ◽  
Mrna And Protein Expression ◽  
Inhibit Cell Proliferation

Background. Previous studies have shown that alkannin has anticancer, anti-inflammatory, and antibacterial effects. However, the effect of alkannin in the development of ovarian cancer (OC) remains unknown. Therefore, this study aims to elucidate the function of alkannin in OC progression. Methods. RT-qPCR and western blot analysis were used to measure mRNA and protein expression. Cell viability and metastasis were detected by the CCK-8 assay, flow cytometry analysis, and transwell assay. Results. Alkannin had no cytotoxicity toward normal ovarian cells, but alkannin can inhibit cell proliferation and induce apoptosis in OC cells. In addition, alkannin inhibited cell migration and invasion and blocked EMT in OC. Besides, upregulation of miR-4461 was found in OC tissues and cells, which was regulated by alkannin. More importantly, miR-4461 can inverse the effects of alkannin on cell viability and metastasis in OC cells. Conclusion. Alkannin restrains cell viability, metastasis, and EMT in OC by downregulating miR-4461 expression.

scholarly journals LncRNA SLC16A1-AS1 Contributes to the Progression of Hepatocellular Carcinoma Cells by Modulating miR-411/MITD1 Axis

2021 ◽  
Author(s):  
Chun Duan ◽  
Bin Quan ◽  
Ni Wang ◽  
Jianghua Yang ◽  
Yan-Lin Yu
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Lines ◽  
Carcinoma Cells ◽  
Luciferase Reporter ◽  
Migration And Invasion ◽  
High Morbidity ◽  
Transwell Assay ◽  
Downstream Target ◽  
Common Malignancy ◽  
Hcc Cells

Abstract Background: Hepatocellular carcinoma (HCC) is a common malignancy with high morbidity. The current study aimed to explore the molecular mechannism of lncRNA SLC16A1-AS1 in the tumorigenesis of HCC.Material and Methods: The expression of SLC16A1-AS1 and miR-411 were examined in clinical HCC tissues. HCC cell lines Hep3B and Huh-7 were employed and transfected with si-SLC16A1-AS1. The correlation between SLC16A1-AS1 and miR-411 was verified by luciferase reporter assay. Cell viability was detected by CCK-8 assay. Cell migration and invasion capacity were examined by transwell assay. The protein level of MITD1 was analyzed by western blotting.Results: The expression of SLC16A1-AS1 markedly increased in HCC tissues and cell lines. Subsequent studies identified SLC16A1-AS1 as a downstream target of miR-411. In addition, SLC16A1-AS1 knockdown and miR-411 overexpression significantly stagnated progression of HCC cells. SLC16A1-AS1 knockdown also downregulated MITD1 levels. Conclusion: Our findings showed that SLC16A1-AS1 was overexpressed in HCC cells and tissues. SLC16A1-AS1 promoted the malignant characteristics of HCC cells and acted as an oncogene. Its regulatory effect may be associated with miR-411/MITD1 axis. Therefore, SLC16A1-AS1 has a potential be used as a biomarker or therapeutic target for the treatment of HCC.

scholarly journals MicroRNA-1271-5p inhibits the tumorigenesis of ovarian cancer through targeting E2F5 and negatively regulates the mTOR signaling pathway

2020 ◽  
Author(s):  
Qin Li ◽  
Junyu Shi ◽  
Xiaoli Xu
Keyword(s):  
Ovarian Cancer ◽  
Signaling Pathway ◽  
Poor Prognosis ◽  
Mtor Signaling ◽  
Luciferase Assay ◽  
Migration And Invasion ◽  
Mtor Signaling Pathway ◽  
Direct Target ◽  
The Relationship

Abstract Background: MicroRNA-1271-5p (miR-1271-5p) has been reported to participate in the progression of many human cancers. However, the role of miR-1271-5p still remains unclear in ovarian cancer (OC). Therefore, we explored the effect of miR-1271-5p on the development of OC in present study. Methods: We measured the miR-1271-5p expression via the qRT-PCR assay. Then the function of miR-1271-5p was analyzed through MTT and Transwell assays. The relationship among miR-1271-5p and E2F5 was verified by dual luciferase assay. The protein expression levels were examined through western blot.Results: MiR-1271-5p was downregulated in OC tissues which predicted poor prognosis of OC patients. Moreover, E2F5 was a direct target of miR-1271-5p in OC. And miR-1271-5p suppressed cell proliferation, migration and invasion in OC through targeting E2F5. Furthermore, E2F5 was upregulated in OC tissues which predicted poor prognosis of OC patients. Besides that, miR-1271-5p suppressed EMT and mTOR pathway in OC. Conclusion: MiR-1271-5p inhibited the tumorigenesis of OC through targeting E2F5 and negatively regulated the mTOR signaling pathway.

scholarly journals MiR-122-5p suppresses the proliferation, migration, and invasion of gastric cancer cells by targeting LYN

2019 ◽  
Vol 52 (1) ◽  
pp. 49-57 ◽  
Author(s):  
Lei Meng ◽  
Zhangming Chen ◽  
Zhe Jiang ◽  
Ting Huang ◽  
Jie Hu ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
Biological Function ◽  
Malignant Tumors ◽  
Luciferase Assay ◽  
Migration And Invasion ◽  
Gastric Cancer Cells ◽  
Mortality And Morbidity ◽  
Qrt Pcr ◽  
The World ◽  
Direct Target

Abstract Gastric cancer (GC) is one of malignant tumors with high mortality and morbidity in the world. MicroRNA-122 (miR-122) acts as a tumor suppressor in a variety of cancers and has been found to be dominant in gastric adenocarcinoma. However, the specific biological function of miR-122-5p in GC is not completely clear. In this study, we found that miR-122-5p was low-expressed in GC tissues and cell lines by using qRT-PCR. Overexpression of miR-122-5p inhibited the proliferation, migration, and invasion of GC cells by using CCK-8 and transwell assays. On the contrary, downregulation of miR-122-5p promoted the proliferation, migration, and invasion of GC cells. In addition, we found that the expression of LYN, an Src family tyrosine kinase, was inversely correlated with miR-122-5p expression in GC tissues by using western blot analysis, immunohistochemistry, and qRT-PCR assays. Meanwhile, luciferase assay results indicated that LYN is a direct target of miR-122-5p in GC cells. Moreover, silencing LYN expression by its siRNA inhibited the proliferation, migration, and invasion of GC cells. Importantly, overexpression of LYN restored miR-122-5p-mediated inhibition of the proliferation, migration, and invasion of GC cells. Taken together, our results indicated miR-122-5p inhibited the proliferation, migration, and invasion by targeting LYN in GC.

scholarly journals Expression of SASH1 in Preeclampsia and Its Effects on Human Trophoblast

2020 ◽  
Vol 2020 ◽  
pp. 1-8
Author(s):  
Shisan Liu ◽  
Sijia Jiang ◽  
Liping Huang ◽  
Yanhong Yu
Keyword(s):  
Flow Cytometry ◽  
Matched Control ◽  
Migration And Invasion ◽  
Flow Cytometry Analysis ◽  
Vector Group ◽  
Transwell Assay ◽  
Human Trophoblast ◽  
Empty Vector ◽  
Group Flow

Aim. To explore the involvement of SASH1 in preeclampsia. Methods. Expression of SASH1 was determined by qPCR, WB, and immunohistochemistry in the placenta of both normal and preeclamptic pregnancies. The SASH1 gene of human HTR-8/SVneo cells was overexpressed by transfection of pEZ-Lv206-SASH1. After that, the CCK-8 assay, EdU assay, transwell assay, and flow cytometry were used to examine the cell proliferation, migration, invasion, and apoptosis. Results. Higher expression of SASH1 was detected in placental tissues collected from patients with preeclampsia, compared with those from gestational age-matched control samples. The expression of SASH1 was significantly enhanced by transfection with pEZ-Lv206-SASH1 in HTR-8/SVneo cells. In addition, the HTR-8/SVneo cells transfected with pEZ-Lv206-SASH1 exhibited significantly reduced proliferation, migration, and invasion ability compared to the cells in the empty vector group and normal group. Flow cytometry analysis demonstrated that the apoptosis rate of cells transfected with pEZ-Lv206-SASH1 was significantly higher than that of cells transfected with empty vector and untreated cells. Conclusions. SASH1 is significantly upregulated in the placenta of preeclampsia, and overexpression of SASH1 can inhibit the proliferation, migration, and invasion, but induce apoptosis of trophoblast cells in vitro.

MiR-18a-5p promotes proliferation, migration and invasion of hepatocellular carcinoma by targeting CPEB3

2020 ◽  
Author(s):  
Jianxing Zheng ◽  
Dongyang Wu ◽  
Libing Wang ◽  
Fengzhi Qu ◽  
Daming Cheng ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Wound Healing ◽  
Expression Profiles ◽  
Bioinformatic Analysis ◽  
Luciferase Assay ◽  
Migration And Invasion ◽  
Promoting Effect ◽  
Migration Ability ◽  
Qrt Pcr ◽  
Hcc Cells

Abstract Objective The study aims to explore the mechanism of miR-18a-5p targeting CPEB3 gene in regulating the occurrence and development of hepatocellular carcinoma (HCC). Methods Differential and survival analyses were conducted on HCC expression profiles from TCGA database to screen out target miRNAs on which targeted prediction was conducted. qRT-PCR was used to detect the expressions of miR-18a-5p and CPEB3. MTT assay examined the proliferation activity, wound healing assay analyzed the migration ability and Transwell assay detected the invasion ability of HCC cells after overexpressing miR-18a-5p.Dual luciferase assay verified the targeting relationship between miR-18a-5p and CPEB3. Meanwhile, MTT, wound healing and Transwellassays determined whether the overexpression of CPEB3 reversed the promoting effect of miR-18a-5p on HCC cells. Results Bioinformatic analysis showed that miR-18a-5p was significantly highly expressed in HCC tissues and its target binding site was found in CPEB3 genewith low expression.The qRT-PCR found that high miR-18a-5p expression was observed in HCC cells, and the expression of CPEB3 was significantly low. Overexpression of miR-18a-5p promoted proliferation, migration and invasion of HCC cells. Dual luciferase assay observed that miR-18a-5p inhibited the expression of CPEB3 while overexpression of CPEB3 reversed the promoting effect of miR-18a-5p on the growth of HCCcells. Conclusion miR-18a-5p promoted the proliferation and migration of HCC cells by inhibiting the expression of CPEB3. The role of miR-18a-5p /CPEB3 in HCCfound in this study provided a new potential target for the prognostic treatment of HCC patients.

scholarly journals MicroRNA-490-3p inhibits migration and chemoresistance of colorectal cancer cells via targeting TNKS2

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Jing Li ◽  
Rubing Mo ◽  
Linmei Zheng
Keyword(s):  
Colorectal Cancer ◽  
Cancer Cells ◽  
Target Genes ◽  
Chemotherapy Resistance ◽  
Luciferase Assay ◽  
Migration And Invasion ◽  
Invasion And Migration ◽  
Colorectal Cancer Cells ◽  
And Migration ◽  
Common Malignancy

Abstract Objective Colorectal cancer is one of the most common malignancy in the world. The oncogenesis of colorectal cancer is still not fully elucidated. It was reported that microRNA-490-3p (miR-490-3p) was closely related to the regulation of cancers. However, if miR-490-3p could also affect colorectal cancer and the specific mechanism remains unclear. Methods qRT-PCR was conducted to examine the expression of miR-490-3p. DIANA, miRDB, and TargetScan databases were used to identify target genes. LOVO and SW480 cells were transfected by miR-490-3p mimics and inhibitors. Transwell assay was used to measure cell invasion and migration. Cisplatin and fluorouracil were administered to investigate chemotherapy resistance. Western blot was used to measure TNKS2 protein expression. Binding sites were verified using the double luciferase assay. Results miR-490-3p expression was low in the colorectal cancer cells. The level of miR-490-3p was negatively correlated with cell migration and invasion of cancer cells. miR-490-3p could bind to TNKS2 mRNA 3′UTR directly. miR-490-3p can suppress cell viability and resistance to chemotherapy in colorectal cancer cells through targeting TNKS2. Conclusions miR-490-3p could affect colorectal cancer by targeting TNKS2. This study may provide a potential therapeutic target for colorectal cancer.

scholarly journals Overexpression of 14-3-3γ Induces the Migration and Invasion of Human Lung Adenocarcinoma A549 Cells

2021 ◽  
Author(s):  
Pritsana Raungrut ◽  
Nidanut Champoochana ◽  
Paramee Thongsuksai ◽  
Kamontip Promnares
Keyword(s):  
Lung Cancer ◽  
Cancer Progression ◽  
A549 Cells ◽  
Migration And Invasion ◽  
P Value ◽  
Transwell Assay ◽  
Scratch Assay ◽  
Invasion And Migration ◽  
Tumor Cell Migration

Objective: 14-3-3 gamma (γ) is known to modulate the development and progression of many cancers. However, the evidence in lung cancer is still unclear. In this study, effects of 14-3-3γ on tumor cell migration and invasion were investigated. Material and Methods: A 14-3-3γ expression vector was made and transfected into A549 cells. In-vitro scratch assay and transwell assay were applied to assess migration and invasion, respectively. Western blotting was used to detect expression of proteins related to epithelial–mesenchymal transition.Results: Closing rate of scratch wounds, both in classical and non-classical scratch assay, was significantly increased in 14-3-3γ-overexpressing cells in comparison to the controls. Similarly, by transwell assay, a significant increase in the invasion and migration was shown in the 14-3-3γ-overexpressing cells in comparison to the null vector cells, by approximately 79.2% (p-value=0.002) and 131.2% (p-value<0.001), respectively. In addition, increased 14-3-3γ expression resulted in a significant increase of β-catenin and Snail but not for E-cadherin and vimentin. Conclusion: The study demonstrates the role of 14-3-3γ protein on lung cancer progression via migration and invasion processes, possibly providing a new targeted therapy for non-small cell lung cancer.

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