Journal: International Journal of Medical Sciences

Article Title: Effect of MT2A on apoptosis and proliferation in HL60 cells

doi: 10.7150/ijms.57821

Figure Lengend Snippet: HL60 cell transfection induces down-regulation of MT2A. (A) FCM analysis of transfection efficiency in the untreated, psiMT2A-1, psiMT2A-2, and psiMT2A-3 group at 72 h post-transfection. (B) qPCR analysis of MT2A expression in each group at mRNA level. GAPDH was used to normalized gene expression. mRNA expression in the psiMT2A-1 group was remarkably down-regulated compared with the control group. (C) Western blot analysis of MT2A down-expression in each group at the protein level. Relative MT2A protein level was quantitatively evaluated by densitometric analysis. The number of independent experiment repetitions was tightly controlled at least three times(right) and represented as mean ± SD. *P < 0.05 vs. control.

Article Snippet: A total of three short hairpin RNA (shRNA) targeted to MT2A mRNA sequence (NM_005953.5), and one negative control vector were designed and synthesized by Shanghai GenePharma Co., LTD. Then an optimal infection sequence was selected in vitro for subsequent experiments.

Techniques: Transfection, Expressing, Gene Expression, Control, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Effect of MT2A on apoptosis and proliferation in HL60 cells

doi: 10.7150/ijms.57821

Figure Lengend Snippet: Transfection of psiMT2A-1 induces HL60 cell proliferation. (A) The proliferation ability of HL60 cells was measured by soft agar colony formation analysis. The number of clones in the psiMT2A-1 group was obviously increased in comparison with the untreated and the NC siRNA group. (B) CCK-8 method was used to evaluate proliferation capacity after silencing of MT2A. (C) Western blot was executed to detect Bax and BCL2 protein expression in the psiMT2A-1, NC siRNA, and untreated groups. The densitometric ratio of Bax/GAPDH and Bcl2/ GAPDH was recorded by quantity one software. Quantification evaluation of data from independent triplicate experiments(right). (D) FCM analysis of apoptosis rates of HL60 cell after MT2A knock-down. The number of independent experiment repetitions was tightly controlled at least three times(right) and represented as mean ± SD. *P < 0.05 vs. control.

Article Snippet: A total of three short hairpin RNA (shRNA) targeted to MT2A mRNA sequence (NM_005953.5), and one negative control vector were designed and synthesized by Shanghai GenePharma Co., LTD. Then an optimal infection sequence was selected in vitro for subsequent experiments.

Techniques: Transfection, Clone Assay, Comparison, CCK-8 Assay, Western Blot, Expressing, Software, Knockdown, Control

Journal: International Journal of Medical Sciences

Article Title: Effect of MT2A on apoptosis and proliferation in HL60 cells

doi: 10.7150/ijms.57821

Figure Lengend Snippet: Detection of MT2A expression in HL60 cells after infection with a lentivirus vector. (A) Assessment of GFP expression in the GV492-MT2A and GV492-KZ group. Magnification, x200. (B) qPCR analysis of MT2A expression in each group at mRNA level after normalization with GAPDH. mRNA expression in the GV492-MT2A group was higher than in control group. (C) Western blot analysis of MT2A over-expression at the protein level. The relative MT2A protein expression level was quantified by densitometric analysis. The number of independent experiment repetitions was tightly controlled at least three times(right) and represented as mean ± SD. * P < 0.05 vs. control.

Article Snippet: A total of three short hairpin RNA (shRNA) targeted to MT2A mRNA sequence (NM_005953.5), and one negative control vector were designed and synthesized by Shanghai GenePharma Co., LTD. Then an optimal infection sequence was selected in vitro for subsequent experiments.

Techniques: Expressing, Infection, Plasmid Preparation, Control, Western Blot, Over Expression

Journal: International Journal of Medical Sciences

Article Title: Effect of MT2A on apoptosis and proliferation in HL60 cells

doi: 10.7150/ijms.57821

Figure Lengend Snippet: MT2A promotes cell apoptosis of HL60 cells. (A) The in vitro soft agar assay of HL60 cells was detected after up-regulation of MT2A. Quantification evaluation of data from independent triplicate experiments(right). (B) Cytotoxicity was examined by the CCK-8 method. The proliferation of GV492-MT2A‑infected HL60 cells was notably suppressed when compared with control. (C) Western blot analysis was used to measure the level of apoptotic proteins Bax and Bcl2. (D) The apoptosis rates of each group were evaluated using FCM analysis. The number of independent experiment repetitions was tightly controlled at least three times (right) and represented as mean ± SD. * P < 0.05 vs. control.

Article Snippet: A total of three short hairpin RNA (shRNA) targeted to MT2A mRNA sequence (NM_005953.5), and one negative control vector were designed and synthesized by Shanghai GenePharma Co., LTD. Then an optimal infection sequence was selected in vitro for subsequent experiments.

Techniques: In Vitro, Soft Agar Assay, CCK-8 Assay, Control, Western Blot

Journal: International Journal of Medical Sciences

Article Title: Effect of MT2A on apoptosis and proliferation in HL60 cells

doi: 10.7150/ijms.57821

Figure Lengend Snippet: Effects of MT2A on cell cycle distribution and NF-κB activity in HL60 human leukemia cells. (A) FCM analysis of HL60 cell cycle changes after MT2A over-expression. Quantification evaluation of data from independent triplicate experiments(right). * P < 0.05 vs. control. (B) Western blot analysis of cyclin- regulated proteins CDK1 and CyclinB1. (1~3: 24,48,72 h after over-expression of MT2A; 4, 72 h after over-expression of MT2A in the GV492-KZ group; 5, Untreated control). (C) Up-regulation of MT2A resulted in increasing protein expression level of IκB-α and decreasing the protein expression level of p-IκB-α and CyclinD1 in HL60 cells.

Article Snippet: A total of three short hairpin RNA (shRNA) targeted to MT2A mRNA sequence (NM_005953.5), and one negative control vector were designed and synthesized by Shanghai GenePharma Co., LTD. Then an optimal infection sequence was selected in vitro for subsequent experiments.

Techniques: Activity Assay, Over Expression, Control, Western Blot, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: Flowchart of the whole work. In a cohort of 191 GC patients and 288 healthy controls, we conducted a questionnaire survey on lifestyle habits and dietary preferences. To obtain genetic information, peripheral blood leukocytes were collected from patients and controls and used for GWAS. After analysis of genetic and nongenetic risk factors, the DHCR7 gene and its mutation site rs104886038 were identified. Further molecular function verification was carried out in vivo and in vitro experiments. Combined with protein structure analysis, we identified and verified the key co-mutation sites of DHCR7, thereby obtaining potential intervention targets for GC treatment

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Mutagenesis, In Vivo, In Vitro

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: rs104886038 and DHCR7 play important roles in GC. A Manhattan plot for the GWAS based on the analysis of the training set with the additive model. B Significant SNPs confirmed in the validation set. C ROC of the SNP model for GC diagnosis. D Forest plots for risk factors for GC. OR(95%CI) and P value of OR were calculated based on the logistic regression model. E In the TCGA and GSE66229 datasets, DHCR7 was upregulated in GC tissues compared with normal tissues (***FDR < 0.001). F The results of functional enrichment analysis of genes coexpressed with DHCR7. G The expression of DHCR7 in GC tissues with different pathological TNM stages, primary tumor invasion depths, lymph node metastasis statuses, distant metastasis statuses and differentiation grades (calculated by Student’s t test, * P < 0.05; ** P < 0.01; *** P < 0.001)

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Functional Assay, Expressing

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: Germline mutations identified by GWAS analysis

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Variant Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: Knockdown of DHCR7 inhibited the proliferation, invasion and migration of GC cells and promoted apoptosis. A Protein expression of DHCR7 was signicicantly upregulated in AGS, MKN-45, MKN-28 and downregulated in HGC-27 cells. B Western blot analysis confirmed that DHCR7 expression was knocked down by siRNAs in GC cell lines. C Proliferation of AGS and MKN-28 cells with different DHCR7 expression levels, as determined by a CCK-8 assay at different time points over a 96 h period. D Results of EdU incorporation assays in AGS and MKN-28 cells transfection with DHCR7 siRNAs. E Results of colony formation assays in AGS and MKN-28 cells transfection with DHCR7 siRNAs. F Results of Transwell migration and invasion assays in AGS and MKN-28 cells transfection with DHCR7 siRNAs. G Results of the apoptosis assay in AGS and MKN-28 cells transfection with DHCR7 siRNAs. The data are presented as the means ± SDs and calculated by Student’s t test. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Migration, Expressing, Western Blot, CCK-8 Assay, Transfection, Apoptosis Assay

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: Overexpression of DHCR7 promotes the proliferation, invasion and migration of GC cells via cellular cholesterol biosynthesis. A Western blot analysis verified that DHCR7 was stably overexpressed in GC cell lines infected with DHCR7 overexpression lentivirus. B Results of EdU incorporation assays in control GC cells, DHCR7-overexpressing GC cells and DHCR7-overexpressing GC cells treated with TAM or AY 9944 for 24 h. C Results of colony formation assays in control GC cells, DHCR7-overexpressing GC cells and DHCR7-overexpressing GC cells treated with TAM or AY 9944 for 14 days. D Results of Transwell migration and invasion assays in control GC cells, DHCR7-overexpressing GC cells and DHCR7-overexpressing GC cells treated with TAM or AY 9944 for 24 h. E Cellular cholesterol level in control GC cells, DHCR7-overexpressing GC cells and DHCR7-overexpressing GC cells treated with TAM or AY 9944 for 24 h, as measured by Filipin III staining. F Cholesterol level in control GC cells, DHCR7-overexpressing GC cells and DHCR7-overexpressing GC cells treated with TAM or AY 9944 for 24 h. AGS were treated with TAM (3 μM) and AY 9944 (5 μM). MKN-28 were treated with TAM (5 μM) and AY 9944 (10 μM). HGC-27 were treated with TAM (5 μM) and AY 9944 (5 μM). The data are presented as the means ± SDs and calculated by Student’s t test. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Over Expression, Migration, Western Blot, Stable Transfection, Infection, Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: Overexpressing DHCR7 increased the cholesterol level and further promoted the malignant phenotype of GC in vivo. A Results of xenograft tumor model generated with DHCR7-overexpressing GC cell and control GC cell ( n = 5 per group). B Representative images of IHC for Ki-67 in tumor tissues from OE-DHCR7 and OE-NC groups. C Results of tissue cholesterol detection in tumors from OE-DHCR7 and OE-NC groups. D Results of murine lung metastasis model generated with DHCR7-overexpressing GC cell and control GC cell ( n = 4 per group). E Representative images of HE stains in lung metastasis from OE-DHCR7 and OE-NC groups. F Results of TAM treatment on DHCR7-overexpressing GC cell in xenograft tumor model ( n = 5 per group). G Representative images of IHC for Ki-67 in tumor tissues from OE-DHCR7 and OE-DHCR7-TAM groups. H Weight of tumors from OE-DHCR7 and OE-DHCR7-TAM groups. I Tissue cholesterol level of tumors from OE-DHCR7 and OE-DHCR7-TAM groups. J Results of TAM treatment on DHCR7-overexpressing GC cell in murine lung metastasis model ( n = 5 per group). K Representative images of HE stains in lung metastasis from OE-DHCR7 and OE-DHCR7-TAM groups. The data represent the means ± SEMs and calculated by Student’s t test. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: In Vivo, Generated

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: Co-mutation of rs104886038 and rs104886035 in DHCR7 reduces its stability and induces its degradation through the ubiquitin–proteasome system. A DHCR7 catalyses the synthesis of cholesterol via both the Bloch and Kandutsch-Russell pathways. B Prediction of the structure of DHCR7. C Amino acid network of DHCR7. D Western blot analysis verified that both wild-type and mutant DHCR7 were overexpressed in sh-DHCR7 GC cells, while the protein expression of mutant DHCR7 was significantly lower than the wild-type. Both wild-type and mutant DHCR7 plasmid were transfected to sh-DHCR7 GC cells. Western blot analysis were performed at 72 h after transfection. E CHX (30 μM) continued to exert effects for 16–24 h, and the level of DHCR7 in the different groups was verified by western blot analysis. F After CHX treatment, WT/MT GC cell lines were treated with CQ (20 μM) and MG132 (10 μM) to verify the degradation pathway of DHCR7. The data are presented as the means ± SDs and calculated by Student’s t test. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Mutagenesis, Western Blot, Expressing, Plasmid Preparation, Transfection

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: The effects of the single L68P mutation and co-mutations with L68P on DHCR7 protein stability

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Mutagenesis

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: Integrated multi-dimensional analysis highlights DHCR7 mutations involving in cholesterol biosynthesis and contributing therapy of gastric cancer

doi: 10.1186/s13046-023-02611-6

Figure Lengend Snippet: Mutant DHCR7 inhibited the proliferation, invasion and migration of GC cells and promoted apoptosis. A The proliferation of GC cells with WT/MT DHCR7 was detected by an EdU incorporation assay. B Results of colony formation assays in GC cells with WT/MT DHCR7. C Results of Transwell migration and invasion assays in GC cells with WT/MT DHCR7. D Results of the apoptosis assay in GC cells with WT/MT DHCR7. E Cholesterol level in GC cells with WT/MT DHCR7. F Cellular cholesterol level, as measured by Filipin III staining. The data are presented as the means ± SDs and calculated by Student’s t test. * P < 0.05; ** P < 0.01; *** P < 0.001

Article Snippet: Lentiviruses carrying DHCR7 cDNA or DHCR7 short hairpin RNA (shRNA) containing the sequence of DHCR7-siRNA-1 were purchased from Shanghai GenePharma Co., Ltd. (Shanghai, China).

Techniques: Mutagenesis, Migration, Apoptosis Assay, Staining

Journal: PLoS ONE

Article Title: Downregulated expression of hepatoma-derived growth factor inhibits migration and invasion of prostate cancer cells by suppressing epithelial-mesenchymal transition and MMP2, MMP9

doi: 10.1371/journal.pone.0190725

Figure Lengend Snippet: (A) Sequence of shRNA-HDGF and shRNA-CTR. (B) Lentivirus transfection efficiency. Fluorescence microscopy (magnification, × 200) demonstrated >80% PCa cells were effectively transfected with shRNA-HDGF and shRNA-CTR lentivirus 24 h after transfection at a MOI of 50 for DU145 and PC3 cells respectively, and 36 h after transfection at a MOI of 20 for LNCaP cells. (C) QRT-PCR analysis revealed that HDGF mRNA levels were down-regulated in DU145, PC3 and LNCaP cells. The relative gene expression level of HDGF was normalized to β-actin. (D, E) Expression of HDGF protein in human PCa cells (DU145, PC3 and LNCaP cells) transfected with lentivirus shRNA-HDGF is reduced compared with the corresponding cells transfected with lentivirus shRNA-CTR or only treated with PBS, and the quantification of HDGF gray intensity of DU145, PC3 and LNCaP cells in each group was presented. The data are presented as mean ± standard deviation of three independent experiments. * P < 0.05 vs PBS, ** P < 0.05 vs shRNA-CTR. PCa, prostate cancer; MOI, multiplicity of infection; HDGF, hepatoma-derived growth factor; shRNA-HDGF, group transfected by recombinant lentivirus shRNA targeting HDGF sequence; shRNA-CTR, group transfected by recombinant lentivirus shRNA with scrambled sequence; PBS, group treated by PBS.

Article Snippet: The recombinant lentivirus short hairpin RNA (shRNA) targeting HDGF sequence (shRNA-HDGF) and control shRNA (shRNA-CTR) were purchased from Bio-Link (Shanghai, China).

Techniques: Sequencing, shRNA, Transfection, Fluorescence, Microscopy, Quantitative RT-PCR, Gene Expression, Expressing, Standard Deviation, Infection, Derivative Assay, Recombinant

Journal: PLoS ONE

Article Title: Downregulated expression of hepatoma-derived growth factor inhibits migration and invasion of prostate cancer cells by suppressing epithelial-mesenchymal transition and MMP2, MMP9

doi: 10.1371/journal.pone.0190725

Figure Lengend Snippet: Scratch assays showed that HDGF knockdown reduced the area of migration of (A, B) DU145 and (C, D) PC3 cells respectively compared with cells transfected by lentivirus shRNA-CTR or treated by PBS. Representative images (magnification, × 200) and quantification of mean area of cell migration of DU145 and PC3 cells in each group are presented. The data are presented as mean ± standard deviation of three independent experiments. * P < 0.05 vs PBS, ** P < 0.05 vs shRNA-CTR. PCa, prostate cancer; HDGF, hepatoma-derived growth factor; shRNA-HDGF, group transfected by recombinant lentivirus shRNA targeting HDGF sequence; shRNA-CTR, group transfected by recombinant lentivirus shRNA with scrambled sequence; PBS, group treated by PBS.

Article Snippet: The recombinant lentivirus short hairpin RNA (shRNA) targeting HDGF sequence (shRNA-HDGF) and control shRNA (shRNA-CTR) were purchased from Bio-Link (Shanghai, China).

Techniques: Knockdown, Migration, Transfection, shRNA, Standard Deviation, Derivative Assay, Recombinant, Sequencing

Journal: PLoS ONE

Article Title: Downregulated expression of hepatoma-derived growth factor inhibits migration and invasion of prostate cancer cells by suppressing epithelial-mesenchymal transition and MMP2, MMP9

doi: 10.1371/journal.pone.0190725

Figure Lengend Snippet: Cell migration capacity of transfected PCa cells by a Transwell assay and a quantification of the number of migrated (A, B) DU145, (C, D) PC3 and (E, F) LNCaP cells in each group. Representative images (magnification, × 200) and a quantification of mean migration number of DU145, PC3 and LNCaP cells in each group are presented. The data are presented as mean ± standard deviation of three independent experiments. * P < 0.05 vs PBS, ** P < 0.05 vs shRNA-CTR. PCa, prostate cancer; HDGF, hepatoma-derived growth factor; shRNA-HDGF, group transfected by recombinant lentivirus shRNA targeting HDGF sequence; shRNA-CTR, group transfected by recombinant lentivirus shRNA with scrambled sequence; PBS, group treated by PBS.

Article Snippet: The recombinant lentivirus short hairpin RNA (shRNA) targeting HDGF sequence (shRNA-HDGF) and control shRNA (shRNA-CTR) were purchased from Bio-Link (Shanghai, China).

Techniques: Migration, Transfection, Transwell Assay, Standard Deviation, shRNA, Derivative Assay, Recombinant, Sequencing

Journal: PLoS ONE

Article Title: Downregulated expression of hepatoma-derived growth factor inhibits migration and invasion of prostate cancer cells by suppressing epithelial-mesenchymal transition and MMP2, MMP9

doi: 10.1371/journal.pone.0190725

Figure Lengend Snippet: Cell invasion capacity of transfected PCa cells by a Transwell assay with Matrigel and a quantification of the number of invaded (A, B) DU145, (C, D) PC3 and (E, F) LNCaP cells in each group. Representative images (magnification, × 200) and quantification of mean invasion number of DU145, PC3 and LNCaP cells in each group are presented. The data are presented as mean ± standard deviation of three independent experiments. * P < 0.05 vs PBS, ** P < 0.05 vs shRNA-CTR. PCa, prostate cancer; HDGF, hepatoma-derived growth factor; shRNA-HDGF, group transfected by recombinant lentivirus shRNA targeting HDGF sequence; shRNA-CTR, group transfected by recombinant lentivirus shRNA with scrambled sequence; PBS, group treated by PBS.

Article Snippet: The recombinant lentivirus short hairpin RNA (shRNA) targeting HDGF sequence (shRNA-HDGF) and control shRNA (shRNA-CTR) were purchased from Bio-Link (Shanghai, China).

Techniques: Transfection, Transwell Assay, Standard Deviation, shRNA, Derivative Assay, Recombinant, Sequencing

Journal: PLoS ONE

Article Title: Downregulated expression of hepatoma-derived growth factor inhibits migration and invasion of prostate cancer cells by suppressing epithelial-mesenchymal transition and MMP2, MMP9

doi: 10.1371/journal.pone.0190725

Figure Lengend Snippet: The relative protein expression levels of Vimentin, N-cadherin, Snail and Slug were reduced, and E-cadherin was increased in shRNA-HDGF transfected (A) DU145, (C) PC3 and (E) LNCaP cells compared with their corresponding control cells transfected with shRNA-CTR or treated with PBS. The quantification of EMT-associated protein gray intensity of (B) DU145, (D) PC3 and (F) LNCaP cells in each group are presented. The data are presented as mean ± standard deviation of three independent experiments. The relative protein expression level was normalized to β-actin. * P < 0.05 vs PBS, ** P < 0.05 vs shRNA-CTR. EMT, epithelial mesenchymal transition; PCa, prostate cancer; HDGF, hepatoma-derived growth factor; shRNA-HDGF, group transfected by recombinant lentivirus shRNA targeting HDGF sequence; shRNA-CTR, group transfected by recombinant lentivirus shRNA with scrambled sequence; PBS, group treated by PBS.

Article Snippet: The recombinant lentivirus short hairpin RNA (shRNA) targeting HDGF sequence (shRNA-HDGF) and control shRNA (shRNA-CTR) were purchased from Bio-Link (Shanghai, China).

Techniques: Expressing, shRNA, Transfection, Control, Standard Deviation, Derivative Assay, Recombinant, Sequencing

Journal: PLoS ONE

Article Title: Downregulated expression of hepatoma-derived growth factor inhibits migration and invasion of prostate cancer cells by suppressing epithelial-mesenchymal transition and MMP2, MMP9

doi: 10.1371/journal.pone.0190725

Figure Lengend Snippet: The relative protein levels of MMP2 and MMP2 were decreased in sh-HDGF transfected (A) DU145, (C) PC3 and (E) LNCaP cells compared with their corresponding control cells transfected with shRNA-CTR or treated with PBS. The gray intensity quantification of MMP2 and MMP9 of (B) DU145, (D) PC3 and (F) LNCaP cells in each group are presented. The data are presented as mean ± standard deviation of three independent experiments. * P < 0.05 vs PBS, ** P < 0.05 vs shRNA-CTR. PCa, prostate cancer; MMP, matrix metalloproteinase; HDGF, hepatoma-derived growth factor; shRNA-HDGF, group transfected by recombinant lentivirus shRNA targeting HDGF sequence; shRNA-CTR, group transfected by recombinant lentivirus shRNA with scrambled sequence; PBS, group treated by PBS.

Article Snippet: The recombinant lentivirus short hairpin RNA (shRNA) targeting HDGF sequence (shRNA-HDGF) and control shRNA (shRNA-CTR) were purchased from Bio-Link (Shanghai, China).

Techniques: Transfection, Control, shRNA, Standard Deviation, Derivative Assay, Recombinant, Sequencing