LncRNA NKX2-1-AS1 As ceRNA Promotes Tumor Progression and Angiogenesis by Upregulating SERPINE1 Expression and Activating VEGFR-2 Signaling Pathway in Gastric Cancer

Abstract Background: Recent evidence indicated that the lncRNA NKX2-1-AS1 (NKX2-1 antisense RNA 1) plays an important role in cancer progression and metastasis. However, the associated molecular mechanisms of NKX2-1-AS1 in GC are still unclear. Methods: To determine the target of the study by bioinformatic analysis. NKX2-1-AS1 expression was measured in paired tumor and non-tumor tissues of 178 GC patients, by quantitative reverse transcription PCR. The in vitro and in vivo biological functions of NKX2-1-AS1 were examined by loss-of-function and gain-of-function experiments. The potential mechanisms of this competing endogenous RNA (ceRNA) were elucidated using dual-luciferase reporter assay, quantitative PCR, Western blot, and fluorescence in situ hybridization (FISH). Results: NKX2-1-AS1 expression was upregulated in GC cell lines and tumor tissues, which was correlated with tumor progression and enhanced angiogenesis. Functionally, NKX2-1-AS1 overexpression promoted GC cell proliferation, metastasis, invasion, and angiogenesis, while NKX2-1-AS1 downregulation reversed these effects, both in vitro and in vivo. Bioinformatics analysis and dual-luciferase assay showed that the microRNA miR-145-5p is a direct target of NKX2-1-AS1, and that NKX2-1-AS1 acts as a ceRNA that regulates angiogenesis in the context of GC. The mechanistic study revealed that miR-145-5p specifically targets serpin family E member 1 (SERPINE1), and that the complex NKX2-1-AS1/miR-145-5p activates VEGFR2 signaling, via SERPINE1, to promote tumor proliferation and angiogenesis. Conclusion: NKX2-1-AS1 upregulation is associated with tumor cell proliferation, increased angiogenesis, and poor prognosis in GC patients. NKX2-1-AS1 regulates SERPINE1 expression and VEGFR2 signaling by acting as a ceRNA for miR-145-5p.

Download Full-text

LncRNA MYLK-AS1 facilitates tumor progression and angiogenesis by targeting miR-424-5p/E2F7 axis and activating VEGFR-2 signaling pathway in hepatocellular carcinoma

10.21203/rs.3.rs-68878/v1 ◽

2020 ◽

Author(s):

Fei Teng ◽

Ju-Xiang Zhang ◽

Qi-Meng Chang ◽

Xu-Bo Wu ◽

Wei-Guo Tang ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Tumor Progression ◽

Underlying Mechanism ◽

Luciferase Reporter ◽

Loss Of Function ◽

Luciferase Reporter Gene ◽

Tumor Tissues ◽

Vegfr2 Signaling

Abstract BackgroundLong non-coding RNA is essential for the metastasis, invasion, angiogenesis and progression of hepatocellular carcinoma (HCC). However, their specific mechanisms are still controversial. Here, we found that Lnc-MYLK-AS1 is a potential oncogene. We systematically analyzed the clinical significance and mechanism of Lnc-MYLK-AS1 in HCC metastasis, invasion and angiogenesis.MethodsDetermine research goals through bioinformatics analysis. The expression of MYLK-AS1 in matched tumor and non-tumor tissues of 156 HCC patients was detected by quantitative reverse transcription PCR. The in vitro and in vivo biological functions of MYLK-AS1 were examined through the loss of function and function gain experiments. The use of dual luciferase reporter gene analysis, quantitative PCR, Western blotting, and fluorescence in situ hybridization (FISH) clarified the underlying mechanism of this competitive endogenous RNA (ceRNA). ResultsMYLK-AS1 is up-regulated in HCC cell lines and tumor tissues, which is related to tumor progression and enhancement of angiogenesis. Overexpression of MYLK-AS1 promotes the proliferation, metastasis, invasion, and angiogenesis of HCC cells, while down-regulation of MYLK-AS1 can reverse these effects in vivo and in vitro. Dual analysis of luciferase and RNA immunoprecipitation showed that microRNA miR-424-5p is the direct target of MYLK-AS1, and MYLK-AS1 acts as ceRNA, which can regulate angiogenesis in HCC. Mechanism studies have shown that miR-424-5p specifically targets E2F transcription factor 7 (E2F7), while the complex MYLK-AS1/miR-424-5p activates VEGFR2 signaling through E2F7, thereby promoting tumor proliferation and angiogenesis.ConclusionThe up-regulation of MYLK-AS1 is related to tumor cell proliferation, increased angiogenesis and poor prognosis in HCC patients. MYLK-AS1 regulates E2F7 expression and VEGFR2 signaling by acting as a ceRNA of miR-424-5p.

Download Full-text

Positive feedback between lncRNA FLVCR1-AS1 and KLF10 may inhibit pancreatic cancer progression via the PTEN/AKT pathway

Journal of Experimental & Clinical Cancer Research ◽

10.1186/s13046-021-02097-0 ◽

2021 ◽

Vol 40 (1) ◽

Author(s):

Jiewei Lin ◽

Shuyu Zhai ◽

Siyi Zou ◽

Zhiwei Xu ◽

Jun Zhang ◽

...

Keyword(s):

Cell Cycle ◽

Pancreatic Cancer ◽

Cell Proliferation ◽

Cancer Progression ◽

Positive Feedback ◽

Molecular Mechanisms ◽

Tumor Tissues ◽

And Migration

Abstract Background FLVCR1-AS1 is a key regulator of cancer progression. However, the biological functions and underlying molecular mechanisms of pancreatic cancer (PC) remain unknown. Methods FLVCR1-AS1 expression levels in 77 PC tissues and matched non-tumor tissues were analyzed by qRT-PCR. Moreover, the role of FLVCR1-AS1 in PC cell proliferation, cell cycle, and migration was verified via functional in vitro and in vivo experiments. Further, the potential competitive endogenous RNA (ceRNA) network between FLVCR1-AS1 and KLF10, as well as FLVCR1-AS1 transcription levels, were investigated. Results FLVCR1-AS1 expression was low in both PC tissues and PC cell lines, and FLVCR1-AS1 downregulation was associated with a worse prognosis in patients with PC. Functional experiments demonstrated that FLVCR1-AS1 overexpression significantly suppressed PC cell proliferation, cell cycle, and migration both in vitro and in vivo. Mechanistic investigations revealed that FLVCR1-AS1 acts as a ceRNA to sequester miR-513c-5p or miR-514b-5p from the sponging KLF10 mRNA, thereby relieving their suppressive effects on KLF10 expression. Additionally, FLVCR1-AS1 was shown to be a direct transcriptional target of KLF10. Conclusions Our research suggests that FLVCR1-AS1 plays a tumor-suppressive role in PC by inhibiting proliferation, cell cycle, and migration through a positive feedback loop with KLF10, thereby providing a novel therapeutic strategy for PC treatment.

Download Full-text

Circ_0001421 facilitates glycolysis and lung cancer development by regulating miR-4677-3p/CDCA3

Diagnostic Pathology ◽

10.1186/s13000-020-01048-1 ◽

2020 ◽

Vol 15 (1) ◽

Author(s):

Koudong Zhang ◽

Hang Hu ◽

Juan Xu ◽

Limin Qiu ◽

Haitao Chen ◽

...

Keyword(s):

Lung Cancer ◽

Cell Proliferation ◽

Tumor Progression ◽

Lactate Production ◽

Tumor Formation ◽

Luciferase Reporter ◽

Circular Rnas ◽

Glucose Assay

Abstract Background Lung cancer (LC) is a malignant tumor originating in the bronchial mucosa or gland of the lung. Circular RNAs (circRNAs) are proved to be key regulators of tumor progression. However, the regulatory effect of circ_0001421 on lung cancer tumorigenesis remains unclear. Methods The expression levels of circ_0001421, microRNA-4677-3p (miR-4677-3p) and cell division cycle associated 3 (CDCA3) were detected by quantitative real-time polymerase chain reaction (qRT-PCR). Methyl thiazolyl tetrazolium (MTT), Transwell and Tumor formation assays were performed to explore the role of circ_0001421 in LC. Glucose consumption and lactate production were examined by a Glucose assay kit and a Lactic Acid assay kit. Western blot was utilized to examine the protein levels of Hexokinase 2 (HK2) and CDCA3. The interaction between miR-4677-3p and circ_0001421 or CDCA3 was confirmed by dual-luciferase reporter assay. Results Circ_0001421 was increased in LC tissues and cells, and knockdown of circ_0001421 repressed cell proliferation, migration, invasion and glycolysis in vitro. Meanwhile, circ_0001421 knockdown inhibited LC tumor growth in vivo. Mechanistically, circ_0001421 could bind to miR-4677-3p, and CDCA3 was a target of miR-4677-3p. Rescue assays manifested that silencing miR-4677-3p or CDCA3 overexpression reversed circ_0001421 knockdown-mediated suppression on cell proliferation, migration, invasion and glycolysis in LC cells. Conclusion Circ_0001421 promoted cell proliferation, migration, invasion and glycolysis in LC by regulating the miR-4677-3p/CDCA3 axis, which providing a new mechanism for LC tumor progression.

Download Full-text

Cyclin D3-CDK6 Complex Facilitates Tumorigenesis by Regulating the C-Myc/miR-15a/16 Axis in a Feedback Loop in Gastric Cancer

10.21203/rs.3.rs-125333/v1 ◽

2020 ◽

Author(s):

Yeting Hong ◽

Wei He ◽

Jianbin Zhang ◽

Lu Shen ◽

Chong Yu ◽

...

Keyword(s):

Gastric Cancer ◽

Cell Cycle ◽

Cell Proliferation ◽

Feedback Loop ◽

Cell Cycle Progression ◽

Molecular Mechanisms ◽

Cyclin D3 ◽

Luciferase Reporter

Abstract Background: Cyclin D3-CDK6 complex is a component of the core cell cycle machinery that regulates cell proliferation. By using Human Protein Atlas database, a higher expression level of this complex was found in gastric cancer. However, the function of this complex in gastric cancer remain poorly understood. This study aims to determine the expression pattern of this complex in gastric cancer and to investigate its biological role during tumorigenesis.Methods: To demonstrate that Cyclin D3-CDK6 regulate the c-Myc/miR-15a/16 axis in a feedback loop in gastric cancer, a series of methods were conducted both in vitro and in vivo experiments, including qRT-PCR, western blot analysis, EdU assay, ﬂow cytometry, luciferase reporter assay and immunohistochemical staining. SPSS and Graphpad prism software were used for data analysis.Results: In this study, we found that Cyclin D3 and CDK6 were significantly upregulated in gastric cancer and correlated with poorer overall survival. Further study proved that this complex significantly promoted cell proliferation and cell cycle progression in vitro and accelerated xenografted tumor growth in vivo. Furthermore, we explored the molecular mechanisms through which the complex mediated Rb phosphorylation and then promoted c-Myc expression in vitro, we also found c-Myc could suppress miR-15a/16 expression in gastric cancer cell. Finally, we found that miR-15a/16 can simultaneously regulate Cyclin D3 and CDK6 expression as direct target genes.Conclusions: Our findings uncover the Cyclin D3-CDK6/c-Myc/miR-15a/16 feedback loop axis as a pivotal role in the regulation of gastric cancer tumorigenesis, and this regulating axis may provide a potential therapeutic target for gastric cancer treatment.

Download Full-text

CircTP63 Promotes Cell Proliferation and Invasion by Regulating EZH2 via Sponging miR-217 in Gallbladder Cancer

10.21203/rs.3.rs-553740/v2 ◽

2021 ◽

Author(s):

Shouhua Wang ◽

Huanjun Tong ◽

Tingting Su ◽

Di Zhou ◽

Weibin Shi ◽

...

Keyword(s):

Cell Proliferation ◽

Tumor Progression ◽

Gallbladder Cancer ◽

In Vivo Studies ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Ezh2 Expression ◽

Proliferation And Invasion

Abstract Background: Gallbladder cancer (GBC) is the most common biliary tract malignancy and has a poor prognosis in patients with GBC. CircRNA TP63 (circTP63) has been implicated in some tumor proliferation and invasion in some tumors. The study aims to investigate the clinical significance and functional role of circTP63 in GBC.Methods: The expression of circTP63 in GBC was detected by qRT-PCR and the association between circTP63 expression and prognosis of GBC patients was analyzed. CCK8 assay, flow cytometry analysis, transwell assay and in vivo studies were used to evaluated the cell proliferation and invasion after circTP63 knockdown in GBC cells. Luciferase reporter assays and RNA pull-down assay were used to determine the correlation between circTP63 and miR-217. Besides, western blot analysis was also performed.Results: In the present study, we showed that circTP63 expression was upregulated in GBC tissues and cells. Higher circTP63 expression was associated with lymph node metastasis and short overall survival (OS) in patients with GBC. In vitro, knockdown of circTP63 inhibited cell proliferation, cell cycle progression, migration and invasion in GBC. Besides, we demonstrated that knockdown of circTP63 inhibited GBC cell EMT process. In vivo, knockdown of circTP63 inhibited tumor growth in GBC. Mechanistically, we demonstrated that circTP63 competitively bind to miR-217 and promoted EZH2 expression and finally facilitated tumor progression.Conclusions: Our findings demonstrated that circTP63 sponge miR-217 and regulated EZH2 expression and finally facilitates tumor progression. Thus, targeting circTP63 may be a therapeutic strategy for the treatment of GBC.

Download Full-text

LncRNA DUXAP8 promotes the progression of laryngeal cancer through targeting miR-384/ POU2F1 axis

10.21203/rs.3.rs-132474/v1 ◽

2020 ◽

Author(s):

Zhanfeng Yan ◽

Xiaohui Wen ◽

Jinsheng Dai ◽

Jinfeng Liu ◽

Pengpeng Hao ◽

...

Keyword(s):

Cell Proliferation ◽

Laryngeal Cancer ◽

Tumor Model ◽

Inhibitory Effect ◽

Luciferase Reporter ◽

Normal Tissues ◽

Tumor Tissues ◽

High Level

Abstract Background Laryngeal cancer is the highest incidence of head and neck cancers in the world. Increasing evidences have demonstrated that long non-coding RNAs (lncRNAs) play crucial roles in the progression of laryngeal cancer. Despite of the essential role of lncRNA DUXAP8 in many human cancers, its function and specific mechanisms in laryngeal cancer are poorly understood. Methods Differentially expression analysis of lncRNAs in GSE59652 dataset was performed by using limma package of R language. The expression of DUXAP8, miR-384 and candidate mRNAs was evaluated by qRT-PCR. Luciferase reporter assay and RIP assay were performed to determine the direct correlation between DUXAP8, miR-384 and POU2F1. Cell proliferation of laryngeal cancer cell lines TU212 and TU177 cells was evaluated by using CCK-8 assay, colony formation assay and EdU staining assay. Xenograft tumor model in vivo and rescue experiments were performed to explore the function and mechanisms of DUXAP8 in laryngeal cancer. Results The expression of DUXAP8 in tumor tissues was higher than that in adjacent normal tissues. High level of DUXAP8 was closely correlated to the worse prognosis of laryngeal cancer patients. Knockdown of DUXAP8 inhibited the proliferation of TU212 and TU177 cells in vitro, as well as tumor growth in vivo. Furthermore, overexpression of POU2F1 significantly attenuated the inhibitory effect of sh-DUXAP8 on cell proliferation of TU212 and TU177 cells. In addition, sh-DUXAP8 significantly decreased the expression of DUXAP8 and POU2F1, while increased miR-384 expression in tumor tissues compared with sh-NC group. Conclusion DUXAP8 acted as a sponge of tumor suppressor miR-384 and then upregulated POU2F1 expression, thereby promoted the development of laryngeal cancer. Our findings suggest that DUXAP8 may serve as a potential therapeutic target for laryngeal cancer.

Download Full-text

miR-362-3p acts as a tumor suppressor by targeting SERBP1 in ovarian cancer

Journal of Ovarian Research ◽

10.1186/s13048-020-00760-2 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Shujun Cao ◽

Na Li ◽

Xihong Liao

Keyword(s):

Ovarian Cancer ◽

Cell Proliferation ◽

Cancer Progression ◽

Target Gene ◽

Molecular Mechanisms ◽

Gynecological Cancer ◽

Luciferase Reporter ◽

Loss Of Function

Abstract Background Ovarian cancer is the leading lethal gynecological cancer and is generally diagnosed during late-stage presentation. In addition, patients with ovarian cancer still face a low 5-year survival rate. Thus, innovative molecular targeting agents are required to overcome this disease. The present study aimed to explore the function of miR-362-3p and the underlying molecular mechanisms influencing ovarian cancer progression. Methods The expression levels of miR-362-3p were determined using qRT-PCR. Gain-of-function and loss-of-function methods were used to detect the effects of miR-362-3p on cell proliferation, cell migration, and tumor metastasis in ovarian cancer. A luciferase reporter assay was performed to confirm the potential target of miR-362-3p, and a rescue experiment was employed to verify the effect of miR-362-3p on ovarian cancer by regulating its target gene. Results miR-362-3p was significantly downregulated in ovarian cancer tissues and cell lines. In vitro, our data showed that miR-362-3p suppressed cell proliferation and migration. In vivo, miR-362-3p inhibited ovarian cancer growth and metastasis. Mechanistically, SERBP1 was identified as a direct target and functional effector of miR-362-3p in ovarian cancer. Moreover, SERBP1 overexpression rescued the biological function of miR-362-3p. Conclusions Our data reveal that miR-362-3p has an inhibitory effect on ovarian cancer. miR-362-3p inhibits the development and progression of ovarian cancer by directly binding its target gene SERBP1.

Download Full-text

Hsa_circ_0026416 promotes proliferation and migration in colorectal cancer via miR-346/NFIB axis

Cancer Cell International ◽

10.1186/s12935-020-01593-1 ◽

2020 ◽

Vol 20 (1) ◽

Author(s):

Yahang Liang ◽

Jingbo Shi ◽

Qingsi He ◽

Guorui Sun ◽

Lei Gao ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Proliferation ◽

Molecular Mechanisms ◽

Xenograft Model ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Potential Biomarker ◽

Mouse Xenograft Model

Abstract Background Colorectal cancer (CRC) is one of the most common cancers worldwide. Circular RNAs (circRNAs), a novel class of non-coding RNAs, have been confirmed to be key regulators of many diseases. With many scholars devoted to studying the biological function and mechanism of circRNAs, their mysterious veil is gradually being revealed. In our research, we explored a new circRNA, hsa_circ_0026416, which was identified as upregulated in CRC with the largest fold change (logFC = 3.70) of the evaluated circRNAs via analysing expression profiling data by high throughput sequencing of members of the GEO dataset (GSE77661) to explore the molecular mechanisms of CRC. Methods qRT-PCR and western blot analysis were utilized to assess the expression of hsa_circ_0026416, miR-346 and Nuclear Factor I/B (NFIB). CCK-8 and transwell assays were utilized to examine cell proliferation, migration and invasion in vitro, respectively. A luciferase reporter assay was used to verify the combination of hsa_circ_0026416, miR-346 and NFIB. A nude mouse xenograft model was also utilized to determine the role of hsa_circ_0026416 in CRC cell growth in vivo. Results Hsa_circ_0026416 was markedly upregulated in CRC patient tissues and plasma and was a poor prognosis in CRC patients. In addition, the area under the curve (AUC) of hsa_circ_0026416 (0.767) was greater than the AUC of CEA (0.670), CA19-9 (0.592) and CA72-4 (0.575). Functionally, hsa_circ_0026416 promotes cell proliferation, migration and invasion both in vitro and in vivo. Mechanistically, hsa_circ_0026416 may function as a ceRNA via competitively absorbing miR-346 to upregulate the expression of NFIB. Conclusions In summary, our findings demonstrate that hsa_circ_0026416 is an oncogene in CRC. Hsa_circ_0026416 promotes the progression of CRC via the miR-346/NFIB axis and may represent a potential biomarker for diagnosis and therapy in CRC.

Download Full-text

CircNOL10 suppresses breast cancer progression by sponging miR-767-5p to regulate SOCS2/JAK/STAT signaling

Journal of Biomedical Science ◽

10.1186/s12929-020-00697-0 ◽

2021 ◽

Vol 28 (1) ◽

Author(s):

Fang Wang ◽

Xiaochun Wang ◽

Jingruo Li ◽

Pengwei Lv ◽

Mingli Han ◽

...

Keyword(s):

Breast Cancer ◽

Cell Proliferation ◽

Cancer Progression ◽

Molecular Mechanisms ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Circular Rnas ◽

Cytokine Signaling

Abstract Background Circular RNAs (circRNAs) have caught increasing attentions and interests for their important involvement in cancer initiation and progression. This study aims to investigate the biological functions of circNOL10 and its potential molecular mechanisms in breast cancer (BC). Materials and methods qRT-PCR and western blot assays were performed to measure the expression of related genes. CCK-8, colony formation, flow cytomerty and transwell assays were used to assess cell proliferation, cell cycle, migration and invasion. RNA pull-down, luciferase reporter and RIP assays were applied to address the potential regulatory mechanism of circNOL10. Results CircNOL10 was down-regulated in BC tissues and cells. Low expression of circNOL10 was associated with larger tumor size, advanced TNM stage, lymph node metastasis and unfavorable prognosis. Overexpression of circNOL10 inhibited cell proliferation, migration, invasion and EMT in vitro and slowed xenograft tumor growth in vivo. Mechanistically, circNOL10 could act as a molecular sponge for miR-767-5p, leading to the up-regulation of suppressors of cytokine signaling 2 (SOCS2) and inactivation of JAK2/STAT5 pathway. Moreover, circNOL10-mediated suppression of malignant phenotypes was attenuated by miR-767-5p. Similar to circNOL10, enforced expression of SOCS2 also resulted in the suppression of cell proliferation and metastasis. Furthermore, knockdown of SOCS2 reversed the tumor-suppressive effect induced by circNOL10. Conclusions CircNOL10 repressed BC development via inactivation of JAK2/STAT5 signaling by regulating miR-767-5p/SOCS2 axis. Our findings offer the possibility of exploiting circNOL10 as a therapeutic and prognostic target for BC patients.

Download Full-text

LncRNA LINC00337 Sponges miR-1285-3p to Promote Proliferation and Metastasis of Lung Adenocarcinoma Cells Through Up-Regulating YTHDF1

10.21203/rs.3.rs-56092/v1 ◽

2020 ◽

Author(s):

Ru-nan Zhang ◽

Dong-mei Wu ◽

Li-ping Wu ◽

Guo-wei Gao

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Lung Adenocarcinoma ◽

Molecular Mechanisms ◽

Luciferase Reporter ◽

Gear Up ◽

Knock Down ◽

Reporter Assays

Abstract Background: Emerging studies have attested that long noncoding RNAs (lncRNAs) predominantly functioned in carcinogenesis of multiple developing human tumors. The current research aimed at probing the underlying participation and mechanisms of LINC00337 in lung adenocarcinoma.Methods: Here we analyzed TCGA and GTEx datasets and chose LINC00337 as research object. Cell proliferation, cell apoptosis, cell cycle, and invasion were detected in gain and loss experiment of LINC00337 both in vitro and in vivo. Moreover, RNA pull-down, luciferase reporter assays, western blotting analysis, rescue experiment were performed to investigate underlying molecular mechanisms of LINC00337 function. Results: LINC00337 was remarkably increased in lung adenocarcinoma. Also, LINC00337 knock-down was unraveled to repress cell invasion and proliferation as well as cell cycle, and gear up apoptosis in lung adenocarcinoma in vitro and in vivo. With respect to mechanism, LINC00337 knock-down boosted miR-1285-3p to be expressed and then restrained YTHDF1 to be expressed post-transcriptionally. Crucially, both miR-1285-3p decrement and YTHDF1 overexpression successfully countered the influence on cell proliferation, invasion and apoptosis caused by LINC00337 shRNA.Conclusions: These results suggest that LINC00337 acted as an oncogenic lncRNA, targeting miR-1285-3p and regulating YTHDF1 expression, to promote the progression of lung adenocarcinoma.

Download Full-text