Effect of Hsa-miRNA-203a-3p on proliferation of skin squamous cell carcinoma-1 human skin squamous cell cancer cells by targeting adenomatous polyposis coli using magnetic nanoparticles

2021 ◽  
Vol 11 (12) ◽  
pp. 1942-1950
Author(s):  
Maojie Cheng ◽  
Dongcheng Gu ◽  
Jurui Feng ◽  
Caixia Li

To investigate the effect of hsa-miR-203a-3p overexpression on the proliferation of human skin squamous cell carcinoma (CSCC) cells and its possible mechanism. Real-time PCR (RT-PCR) was used to detect the expression of miR-203a-3p in cell lines and clinical human CSCC samples. A luciferase reporter system was used to verify the targeted regulatory relationship of miR-203a-3p to APC, and a miR-203a-3p lentivirus overexpression vector was constructed and used to transfect CSCC SCL-1 cells. RT-PCR was used to detect changes in miR-203a-3p and APC gene expression and Western blot was used to detect differences in APC and β-catenin protein expression. MTT and clonogenic assays were used to evaluate cell growth and detect clone formation, respectively. MiR-203a-3p showed decreased expression in SCL-1 cells and CSCC samples. Results of luciferase reporter assay showed that the ratio of Renilla luciferase to Firefly luciferase was significantly decreased in SCL-1 cells of the APC 3′-UTR+miR-203a-3p (wild-type) group compared with those of the APC 3′-UTR+negative control group. After lentiviral infection of SCL-1 cells, the abundance of miR-203a-3p and phosphorylated β-catenin protein was significantly increased, whereas the abundance of APC and β-catenin protein was significantly reduced. Cell phenotyping analysis showed that miR-203a-3p decreased cell proliferation. MiR-203a-3p inhibits the proliferation of SCL-1 cells through targeted regulation of APC and may play a role as a tumor suppressor gene through the Wnt pathway. Nanotechnology has potential future research applications in gene vector transfection technology.

Author(s):  
Zheng Dong ◽  
Qing-Hua Xu ◽  
Yuan-Bin Zhu ◽  
Yong-Feng Wang ◽  
Jie Xiong ◽  
...  

Aims : The present study explored the clinical significance of microRNA-22 (miR-22) expression in lung squamous cell carcinoma and to explore the targeting relationship with vascular endothelial growth factor receptor 3 (VEGFR3). Methods: A total of 49 patients with lung squamous cell carcinoma who underwent surgical treatment was selected. The expression of miR-22 was detected by fluorescence quantitative real-time PCR (qPCR), the expression of VEGFR3 was detected by Western blotting assays, and D240 labeled microlymphatic vessels density (MLVD) was detected immunohistochemistry (IHC). Lung squamous cell carcinoma cell line SK-MES-1 was selected and the targeting relationship between miR-22 and VEGFR3 was analyzed by double luciferase reporter gene assay. Western blotting assays were used to detect the expression of vascular endothelial growth factor-D (VEGF-D) and D240 in the blank control group, empty vector transfection group, miR-22 transfection group, miR-22 and VEGFR3 co-transfection group. Results: The expression range of miR-22 in lung squamous cell carcinoma was 0.8-3.5. The expression of miR-22 in lung squamous cell carcinoma was significantly different by tumor maximum diameter, lymph node metastasis, vascular invasion and TNM stage. The expression of miR-22 was linked to survival time. There was a negative correlation between miR-22 and VEGFR3, miR-22 and MLVD. Double luciferase reporter gene assays showed that miR-22 reduced the luciferase activity of pGL3-VEGFR3-WT transfected cells. Compared with the control group, the expression of VEGF-D and D2-40 in the miR-22 transfection group was significantly decreased. However, VEGF-D and D240 in the miR-22 and VEGFR3 cotransfection group reversed the changes. Conclusion: We assumed that the abnormal expression of miR-22 in lung squamous cell carcinoma may be involved in the development and progression of lung squamous cell carcinoma. MiR-22 negatively regulated the target gene VEGFR3 to mediate lymphangiogenesis. The expression of miR-22 may also be linked to the prognosis of the disease.


2018 ◽  
Vol 49 (4) ◽  
pp. 1329-1341 ◽  
Author(s):  
Nan Li ◽  
Chuan-Chuan Nan ◽  
Xue-Yun Zhong ◽  
Jun-Quan Weng ◽  
Hai-Dong Fan ◽  
...  

Background/Aims: Emerging evidence suggests that the propagation of oral squamous cell carcinoma (OSCC) is influenced by the abnormal expression of microRNAs (miRNAs). This study aimed to characterize the involvement of miR-182-5p in OSCC by targeting the calcium/ calmodulin-dependent protein kinase II inhibitor CAMK2N1. Methods: miR-182-5p expression was quantified in OSCC tissues and cell lines with reverse transcription polymerase chain reaction (RT-PCR). Cell colony formation, Cell Counting Kit-8 (CCK-8), Ki-67, and nude mouse xenograft assays were used to characterize the role of miR-182-5p in the proliferation of OSCC. A miR-182-5p target gene was identified with western blotting, RT-PCR, and luciferase activity assays. OSCC patient survival based on CAMK2N1 expression was also analyzed. Results: miR-182-5p was up-regulated in in vitro cell lines and in vivo clinical OSCC samples. CCK-8, colony formation, and Ki-67 assays revealed that miR-182-5p promoted the growth and proliferation of OSCC cells. miR-182-5p directly targeted CAMK2N1, as evidenced by luciferase assays and target prediction algorithms. CAMK2N1 operated as a tumor suppressor gene in patients with OSCC. Down-regulating miR-182-5p expression in the CAL-27 cell line restored CAMK2N1-mediated OSCC cell proliferation. miR-182-5p expression inhibited the activation of AKT, ERK1/2, and NF-κB. Mice injected with CAL-27 cells transfected with miR-182-5p-inhibitor demonstrated a significant increase in tumor size and weight and increased CAMK2N1 mRNA and protein expression compared with the miR-negative control group. Conclusion: The miR-182-5p-CAMK2N1 pathway can be potentially targeted to regulate the proliferation of OSCC cells.


Fitoterapia ◽  
2013 ◽  
Vol 86 ◽  
pp. 84-91 ◽  
Author(s):  
Yuqin Hao ◽  
Weixing Huang ◽  
Mingmei Liao ◽  
Yude Zhu ◽  
Hong Liu ◽  
...  

Oncotarget ◽  
2021 ◽  
Vol 12 (7) ◽  
pp. 638-648
Author(s):  
Julien Schaller ◽  
Hélène Maby-El Hajjami ◽  
Sylvie Rusakiewicz ◽  
Kalliopi Ioannidou ◽  
Nathalie Piazzon ◽  
...  

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