Journal: Cancers

Article Title: MDK Activates the PI3K/AKT Axis to Induce AP2A1 Expression and Epithelial–Mesenchymal Transition in Colorectal Cancer

doi: 10.3390/cancers18081311

Figure Lengend Snippet: Correlation between MDK and AP2A1 and the expression of AP2A1 in CRC tissues and cells. ( A ) Volcano plot of RNA-seq data showing genes differentially expressed upon MDK knockdown (sh-MDK vs. sh-NC). Down- and up-regulated genes are highlighted in blue and red, respectively. ( B , C ) Correlation analyses between MDK and AP2A1 expression in the TCGA ( B ) and LinkedOmics ( C ) CRC cohorts. ( D , E ) Validation of MDK (( D ), GSE81558 ) and AP2A1 (( E ), TCGA) upregulation in CRC tumors compared to normal tissues. ( F – I ) MDK regulates AP2A1 expression. AP2A1 mRNA ( F , G ) and protein ( H , I ) levels were assessed by qRT-PCR and immunoblotting in cells with MDK overexpression or knockdown. Data are presented as mean ± SD; * p < 0.05, (Student’s t -test). The uncropped blots are shown in .

Article Snippet: Commercially synthesized human MDK and AP2A1 siRNAs were purchased from Shanghai Generay Biotech Co., Ltd. (Shanghai, China), with detailed information listed in .

Techniques: Expressing, RNA Sequencing, Knockdown, Biomarker Discovery, Quantitative RT-PCR, Western Blot, Over Expression

Journal: Cancers

Article Title: MDK Activates the PI3K/AKT Axis to Induce AP2A1 Expression and Epithelial–Mesenchymal Transition in Colorectal Cancer

doi: 10.3390/cancers18081311

Figure Lengend Snippet: AP2A1 functions as a downstream regulator of MDK-driven CRC invasion. ( A , B ) Immunoblotting verified that altering MDK levels correspondingly changes AP2A1 expression. Band intensities were quantified and normalized to Tubulin. ( C – F ) Functional rescue experiments showed that the increased invasion/migration caused by MDK overexpression (or the decrease caused by its knockdown) was reversed by co-manipulating AP2A1, as demonstrated by Transwell assays (with or without Matrigel). ×100 magnification; scale bar not shown. ( G , H ) Likewise, MDK-induced cell growth phenotypes were affected by changing AP2A1 levels. ×20 magnification; scale bar not shown. * p < 0.05 vs. control; # p < 0.05 vs. MDK-manipulated group (Student’s t -test). The uncropped blots are shown in .

Article Snippet: Commercially synthesized human MDK and AP2A1 siRNAs were purchased from Shanghai Generay Biotech Co., Ltd. (Shanghai, China), with detailed information listed in .

Techniques: Western Blot, Expressing, Functional Assay, Migration, Over Expression, Knockdown, Control

Journal: Cancers

Article Title: MDK Activates the PI3K/AKT Axis to Induce AP2A1 Expression and Epithelial–Mesenchymal Transition in Colorectal Cancer

doi: 10.3390/cancers18081311

Figure Lengend Snippet: MDK knockdown inhibits tumor growth in vivo. ( A ) IHC analysis of xenograft tumors for the expression of Ki-67, β-catenin, MMP2, MMP9, N-cadherin, and Vimentin in the control (NC) and MDK-KD groups. Scale bars = 200 μm. All data are presented as mean ± SD. ( B ) Western blot analysis of MDK and AP2A1 expression in the tumors. ( C ) qRT-PCR analysis of MDK expression in the tumors. Data are shown as means ± SD. ** p < 0.01. The uncropped blots are shown in .

Article Snippet: Commercially synthesized human MDK and AP2A1 siRNAs were purchased from Shanghai Generay Biotech Co., Ltd. (Shanghai, China), with detailed information listed in .

Techniques: Knockdown, In Vivo, Expressing, Control, Western Blot, Quantitative RT-PCR

Journal: Cancers

Article Title: MDK Activates the PI3K/AKT Axis to Induce AP2A1 Expression and Epithelial–Mesenchymal Transition in Colorectal Cancer

doi: 10.3390/cancers18081311

Figure Lengend Snippet: MDK-regulated AP2A1 expression and cell invasion depend on the activation of PI3K/AKT signaling. The expression of EMT markers ( A , B ) and key components of the PI3K/AKT pathway ( C ) was examined in CRC cells after co-transfection to manipulate MDK and AP2A1 levels. ( D ) The role of PI3K/AKT signaling was confirmed using the inhibitor LY294002. SW480 and RKO cells were pre-treated with LY294002 48 h post-transfection, then assessed for AP2A1 expression. Band intensities were quantified and normalized to GAPDH. ( E – G ) Functional effects of pathway inhibition: After LY294002 pre-treatment, the invasion, migration, and growth of SW480 cells were evaluated. ( E , F ): ×100 magnification; scale bar not shown. ( G ): ×20 magnification; scale bar not shown. * p < 0.05 vs. control; # p < 0.05 vs. MDK-overexpressing cells (Student’s t -test). The uncropped blots are shown in .

Article Snippet: Commercially synthesized human MDK and AP2A1 siRNAs were purchased from Shanghai Generay Biotech Co., Ltd. (Shanghai, China), with detailed information listed in .

Techniques: Expressing, Activation Assay, Cotransfection, Transfection, Functional Assay, Inhibition, Migration, Control

Journal: Cancers

Article Title: MDK Activates the PI3K/AKT Axis to Induce AP2A1 Expression and Epithelial–Mesenchymal Transition in Colorectal Cancer

doi: 10.3390/cancers18081311

Figure Lengend Snippet: Schematic illustration of the mechanism by which MDK promotes EMT progression and induces AP2A1 levels in CRC cells.

Article Snippet: Commercially synthesized human MDK and AP2A1 siRNAs were purchased from Shanghai Generay Biotech Co., Ltd. (Shanghai, China), with detailed information listed in .

Techniques: