Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Expression of CD147 and HE4 in different ovarian tissues

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Relationships between the expression of CD147, HE4 and clinicopathological parameters of 96 ovarian cancer patients

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing, Significance Assay, Biomarker Discovery

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Expression and co-immunoprecipitation of CD147 and HE4 in ovarian cancer cells. (A) The expression of CD147 and HE4 in ovarian cancer cell lines (CaoV3, OVCAR3, SKOV3 and ES-2) detected by Western blot. (B) Cell lysates from OVCAR3 and ES-2 cells were subjected to immunoprecipitation with anti-HE4 antibody and immunoblotted with anti-CD147 antibody. (C) Cell lysates from OVCAR3 and ES-2 cells was immunoprecipitated with anti-CD147 antibody and immunoblotted with anti-HE4 antibody, “IgG” represents the negative control. “Input” indicates total cell lysate of OVCAR3 and ES-2.

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing, Immunoprecipitation, Western Blot, Negative Control

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Co-localization of CD147 and HE4 in ovarian cancer cells. (A) Expression of CD147 and HE4 in ovarian cancer cell lines OVCAR3 (a, d), ES-2 (b, e), SKOV3 (c, f) conducted by immunocytochemistry. (B) The co-localization of CD147 (green) and HE4 (red) in OVCAR3 (a-d) and ES-2 cells (e-h) analyzed by double-labeling immunofluorescence assays, "blue" represents nucleus; "orange" represents the co-localization of CD147 and HE4. The small box at the top-left corner was magnified by 400× (A, B).

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing, Immunocytochemistry, Labeling, Immunofluorescence

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Expression and co-localization of CD147 and HE4 in different ovarian tissues. (A) The expression of CD147 and HE4 in malignant group (a, e), borderline group (b, f), benign group (c, g), and normal ovarian tissues (d, h) detected by immunohistochemistry. (B) Co-localization of CD147 and HE4 analysed by double-labeling immunofluorescence assay in malignant group (a-d), borderline group (e-h), benign group (i-l), and normal tissues (m-p). The color of “green” indicates CD147; “red” indicates HE4; “blue” indicates nucleus; “orange” indicates the co-localization of CD147 and HE4. The small box at the top-left corner was magnified by 400× (A, B).

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing, Immunohistochemistry, Labeling, Immunofluorescence

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Relationship between CD147, HE4 expression and clinicopathologic parameters. (A, D) Compared the expression of CD147, HE4 protein in FIGO stages (I-II) with stages (III-IV) in OC, respectively; (B, C) Compared expression of CD147, HE4 protein in well differentiation group with poor differentiation group in OC, respectively. (E) Compared expression of HE4 protein in lymph node metastasis with non-metastasis group in OC. OC: ovarian cancer.

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Kaplan-Meier analysis of the prognosis of ovarian cancer. Relationship between prognosis and high or low CD147 expression ( A ), high or low HE4 expression ( B ), FIGO stages ( C ), lymphnode metastasis ( D ).

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: The linear regression analysis of CD147 and HE4 expression in ovarian cancer

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing, Biomarker Discovery

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: The correlation between the expression of CD147 and HE4 in ovarian cancer (the Spearman correlation coefficient Rs=0.708, P =0.0000)

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Interaction between CD147 and HE4 expression. (A) The expression of HE4 protein decreased after downregulation of CD147 protein in OVCAR3 and ES-2 cells by Western blot. (B) The expression of CD147 was downregulated after inhibition of HE4 protein in OVCAR3 and ES-2 cells by Western blot. OV: OVCAR3, ES: ES-2, HE4: human epididymis protein 4, OV-NC: negative control group of OVCAR3 cells, ES-NC: negative control of ES-2 cells, OV-CD147-L: downregulation of CD147 expression in OVCAR3 constructed by lentivirus, ES-2-CD147-L: downregulation of CD147 expression in ES-2 constructed by lentivirus, OV-siHE4-L-1/2: downregulation of HE4 expression in OVCAR3 by siRNA, ES-siHE4-L-1/2: inhibition of HE4 expression in ES-2 by siRNA.

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Expressing, Western Blot, Inhibition, Negative Control, Construct

Journal: Journal of Cancer

Article Title: Interaction of CD147 and human epididymis protein 4 promotes invasion and metastasis of ovarian cancer

doi: 10.7150/jca.62440

Figure Lengend Snippet: Interaction of CD147 and HE4 promotes invasion and migration of ovarian cancer cells. (A, C) The invasion capacities of ovarian cancer cells (OVCAR3 and ES-2) after downregulation of CD147 protein and addition of recombinant HE4 active protein detected by Transwell assay. (B, D) The migration capacities of ovarian cancer cells (OVCAR3 and ES-2) after downregulation of CD147 protein and addition of recombinant HE4 active protein detected by Scratch test. HE4: human epididymis protein 4.

Article Snippet: The immunoprecipitation was analyzed by 10%SDS gel electrophoresis, and rabbit anti-HE4 monoclonal antibody (Abcam) and rabbit anti-CD147 polyclonal antibody (Proteintech) were used for western blot analysis.

Techniques: Migration, Recombinant, Transwell Assay

Journal: International Journal of Molecular Sciences

Article Title: Promoter Methylation of Cancer Stem Cell Surface Markers as an Epigenetic Biomarker for Prognosis of Oral Squamous Cell Carcinoma

doi: 10.3390/ijms232314624

Figure Lengend Snippet: Epigenetic inactivation of CSC surface markers ( CD24 , CD44 , CD133 , and CD147 ) in OSCC cell lines. Quantitative real-time RT-PCR analysis was carried out to assess the transcriptional expression level of ( A ) CD24 , ( B ) CD44 , ( C ) CD133 , and ( D ) CD147 genes in OSCC cell lines before and after treatment with 5-aza-dC (5 μM) for 72 h and TSA (0.3 μM) for 18 h. The expression levels of the genes were internally normalized to the expression levels of GAPDH , and the normalized expression for each gene before 5-aza-dC treatments was set to one. The asterisks indicate significant increases in gene expression after 5-aza-dC treatment ( p < 0.05). All data were statistically analyzed using the Student’s t -test. * p < 0.05; ** p < 0.01; *** p < 0.001; no asterisk, not significant. A two-tailed Student’s t -test was used to compare data between the two groups.

Article Snippet: Primary antibodies against CD133 (1:100; #66666-1) (Proteintech, Rosemont, IL, USA) and CD147 (1:200; #13287) (Cell Signaling Technology, Danvers, MA, USA) with a benchmark autostainer (Roche Tissue Diagnostics, Oro Valley, AZ, USA) were used following the manufacturer’s protocol.

Techniques: Quantitative RT-PCR, Expressing, Gene Expression, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Promoter Methylation of Cancer Stem Cell Surface Markers as an Epigenetic Biomarker for Prognosis of Oral Squamous Cell Carcinoma

doi: 10.3390/ijms232314624

Figure Lengend Snippet: Promoter DNA methylation analysis of the CD24 , CD44 , CD133 , and CD147 genes in OSCC cell lines, primary OSCC tumor tissues, and normal oral mucosal tissues. ( A ) Schematic structure of the CpG islands at the promoter regions for CD24 , CD44 , CD133 , and CD147. ( A ) The primer locations for MSP were indicated as an orange bar for each gene. ( B ) MSP results in OSCC cell lines. Gel pictures describe the methylation analysis by MSP. DNA methyltransferase 1 and 3b knockout HCT116 cells (DKO) were included as positive controls. The PCR products recognized the unmethylated (U) and methylated (M) genes. DKO cells were used for the unmethylated control. IVD = in vitro methylated control; ddH 2 O = water control containing no DNA. ( C ) Methylation frequency of the CD24 , CD133 , and CD147 genes between the samples from patients with OSCC ( n = 102) and the normal oral mucosal tissue samples ( n = 45).

Article Snippet: Primary antibodies against CD133 (1:100; #66666-1) (Proteintech, Rosemont, IL, USA) and CD147 (1:200; #13287) (Cell Signaling Technology, Danvers, MA, USA) with a benchmark autostainer (Roche Tissue Diagnostics, Oro Valley, AZ, USA) were used following the manufacturer’s protocol.

Techniques: DNA Methylation Assay, Methylation, Knock-Out, Control, In Vitro

Journal: International Journal of Molecular Sciences

Article Title: Promoter Methylation of Cancer Stem Cell Surface Markers as an Epigenetic Biomarker for Prognosis of Oral Squamous Cell Carcinoma

doi: 10.3390/ijms232314624

Figure Lengend Snippet: Representative bisulfite sequencing results of the CpG islands in the CD24 , CD133 , and CD147 gene promoter regions in OSCC cell lines, primary OSCC tumors ( n = 3), and normal oral mucosal tissues ( n = 3). The locations of the CpG sites ( CD24 : upstream region from −903 to −770; CD133: exon 1 region from −230 to +90 ; and CD147: upstream region from −552 to −291) relative to the transcription start sites (TSSs) of exon 1 are shown. Each box represents a CpG dinucleotide. The black boxes represent methylated cytosines, and the white boxes represent unmethylated cytosines.

Article Snippet: Primary antibodies against CD133 (1:100; #66666-1) (Proteintech, Rosemont, IL, USA) and CD147 (1:200; #13287) (Cell Signaling Technology, Danvers, MA, USA) with a benchmark autostainer (Roche Tissue Diagnostics, Oro Valley, AZ, USA) were used following the manufacturer’s protocol.

Techniques: Methylation Sequencing, Methylation

Journal: International Journal of Molecular Sciences

Article Title: Promoter Methylation of Cancer Stem Cell Surface Markers as an Epigenetic Biomarker for Prognosis of Oral Squamous Cell Carcinoma

doi: 10.3390/ijms232314624

Figure Lengend Snippet: Correlation analysis between gene expression level and promoter methylation level from the TCGA database. ( A , D ) The expression levels of CD133 (PROM1) and CD147 (BSG) in tumor tissues were inversely correlated with the promoter DNA methylation rates. Expression levels of ( A ) CD133 and ( D ) CD147 in primary OSCC tumors ( n = 328) compared to normal oral mucosa ( n = 32), extracted from the TCGA HNSCC database. ( B , E ) The methylation levels of the CpG sites of ( B ) CD133 and ( E ) CD147 in primary OSCC tumors ( n = 338) compared to normal oral mucosa ( n = 34), extracted from the HNSCC TCGA database. ( C , F ) Negative correlation between the gene expression and promoter methylation of ( C , F ). Spearman’s correlation analysis was performed between the methylation (vertical axis) and gene expression (horizontal axis) of the CD133 ( R = −0.20, p = 0.002686) and CD147 genes ( R = −0.17, p = 0.0002601). The Spearman’s correlation coefficients and p -values are shown in each plot.

Article Snippet: Primary antibodies against CD133 (1:100; #66666-1) (Proteintech, Rosemont, IL, USA) and CD147 (1:200; #13287) (Cell Signaling Technology, Danvers, MA, USA) with a benchmark autostainer (Roche Tissue Diagnostics, Oro Valley, AZ, USA) were used following the manufacturer’s protocol.

Techniques: Gene Expression, Methylation, Expressing, DNA Methylation Assay

Journal: International Journal of Molecular Sciences

Article Title: Promoter Methylation of Cancer Stem Cell Surface Markers as an Epigenetic Biomarker for Prognosis of Oral Squamous Cell Carcinoma

doi: 10.3390/ijms232314624

Figure Lengend Snippet: Protein expression levels of CD133 and CD147 in primary OSCC tumors and normal oral mucosal tissue samples. The representative immunohistochemical analysis results show ( A ) CD133 and ( B ) CD147 expression in primary OSCC tissues and normal oral mucosal tissues (left images: × 40, scale bar, 50 µm; right images: × 20, scale bar, 100 µm).

Article Snippet: Primary antibodies against CD133 (1:100; #66666-1) (Proteintech, Rosemont, IL, USA) and CD147 (1:200; #13287) (Cell Signaling Technology, Danvers, MA, USA) with a benchmark autostainer (Roche Tissue Diagnostics, Oro Valley, AZ, USA) were used following the manufacturer’s protocol.

Techniques: Expressing, Immunohistochemical staining

Journal: International Journal of Molecular Sciences

Article Title: Promoter Methylation of Cancer Stem Cell Surface Markers as an Epigenetic Biomarker for Prognosis of Oral Squamous Cell Carcinoma

doi: 10.3390/ijms232314624

Figure Lengend Snippet: Kaplan-Meier curves showing the effect of CD133 and CD147 DNA methylation on overall survival among patients with OSCC ( n = 338) from the HNSCC TCGA dataset. The overall survival for ( A ) CD133 ( p = 0.018) and ( B ) CD147 ( p = 0.0054), respectively, is shown. A probability of <0.05 was considered to represent a statistically significant difference. ( C ) Combination of CD133 and CD147 gene methylation ( p = 0.025). The clinical information of patients with OSCC ( n = 338) was divided into two groups according to their methylation status (β-values) on CD133 (methylation high (red line), n = 209 and methylation low (blue line), n = 129), CD147 (methylation high (red line), n = 113 and methylation low (blue line), n = 55), and a combination of CD133 and CD147 (methylation high (red line), n = 143 and methylation low (blue line), n = 195).

Article Snippet: Primary antibodies against CD133 (1:100; #66666-1) (Proteintech, Rosemont, IL, USA) and CD147 (1:200; #13287) (Cell Signaling Technology, Danvers, MA, USA) with a benchmark autostainer (Roche Tissue Diagnostics, Oro Valley, AZ, USA) were used following the manufacturer’s protocol.

Techniques: DNA Methylation Assay, Methylation

Journal: Cell stress & chaperones

Article Title: Heat shock protein A12A activates migration of hepatocellular carcinoma cells in a monocarboxylate transporter 4-dependent manner.

doi: 10.1007/s12192-021-01251-z

Figure Lengend Snippet: Fig. 5 HSPA12A formed complex with MCT4 in HCC cells. a Mass spectrometry. HepG2 cells were overexpressed with the flag-tagged HSPA12A (Ad-HSPA12A) or control flag adenovirus (Ad-NC). Cel- lular protein extracts were immunoprecipitated with primary antibody for flag. The immunoprecipitates were subjected to mass spectro- metric analysis. The MS/MS and main sequence ions of representa- tive MCT4 peptide EPQPEVAAAEEEK + 2 (m/z: 713.8378) and LLDLSVFR + 2 (m/z: 401.7570) were shown. b Comparison of pep- tide count of MCT4 by mass spectrometry. All data were presented as mean ± SD. *P < 0.05, n = 3/group. c Immunoprecipitation-immunob- lotting analysis. HepG2 cells were overexpressed with HSPA12A by

Article Snippet: Primary antibody against CD147, PKM2, and MCT4 were from Proteintech Group (Rosemont, IL).

Techniques: Mass Spectrometry, Control, Immunoprecipitation, Tandem Mass Spectroscopy, Sequencing, Comparison

Journal: Cell stress & chaperones

Article Title: Heat shock protein A12A activates migration of hepatocellular carcinoma cells in a monocarboxylate transporter 4-dependent manner.

doi: 10.1007/s12192-021-01251-z

Figure Lengend Snippet: Fig. 6 HSPA12A increased MCT4 expression and cell membrane localization of HCC cells. HepG2 cells were infected with Hspa12a- adenovirus to overexpress HSPA12A (Ad-HSPA12A) and HepG2 cells infected with empty virus served as negative controls (Ad- NC). Seventy-two hours after infection, the following measurements were performed. a MCT4 mRNA expression. The mRNA levels of MCT4 in HepG2 cells were determined by quantitative PCR. The PCR results of GAPDH served as internal controls. All data were pre- sented as mean ± SD. *P < 0.05, n = 5/group. b MCT4 protein expres- sion. The protein levels of MCT4 in HepG2 cells were evaluated by

Article Snippet: Primary antibody against CD147, PKM2, and MCT4 were from Proteintech Group (Rosemont, IL).

Techniques: Expressing, Membrane, Infection, Virus, Real-time Polymerase Chain Reaction

Journal: Cell stress & chaperones

Article Title: Heat shock protein A12A activates migration of hepatocellular carcinoma cells in a monocarboxylate transporter 4-dependent manner.

doi: 10.1007/s12192-021-01251-z

Figure Lengend Snippet: Fig. 7 Knockdown of MCT4 mitigated the HSPA12A-induced lac- tate output and migration of HCC cells. HepG2 cells were infected with Hspa12a-adenovirus to overexpress HSPA12A (Ad-HSPA12A). Subsequently, the Ad-HSPA12A HepG2 cells were transfected with MCT4-targeted siRNA (Si-MCT4) to knockdown MCT4 expression or transfected with scramble control siRNA (Si-NC). Forty-eight hours after Si-RNA transfection, the following measurements were performed. a MCT4 expression. Expression of MCT4 was examined by immunoblotting. The blots against α-tubulin served as loading controls. All data were presented as mean ± SD. **P < 0.01, n = 3/ group. b Extracelluar lactate contents. Measurement of extracelluar lactate levels was performed. All data were presented as mean ± SD. *P < 0.05, n = 3/group. c Wound-healing assay. Representative images showed wound healing of Ad-NC HepG2 cells, Si-NC and Si-MCT4

Article Snippet: Primary antibody against CD147, PKM2, and MCT4 were from Proteintech Group (Rosemont, IL).

Techniques: Knockdown, Migration, Infection, Transfection, Expressing, Control, Western Blot, Wound Healing Assay

Journal: Cell stress & chaperones

Article Title: Heat shock protein A12A activates migration of hepatocellular carcinoma cells in a monocarboxylate transporter 4-dependent manner.

doi: 10.1007/s12192-021-01251-z

Figure Lengend Snippet: Fig. 8 Mechanistic scheme. HSPA12A upregulates MCT4 expression, interacts with MCT4, as well as increases MCT4 localization on cell membrane, thereby facilitates the efflux of lactate that gener- ated from glycolysis, and ulti- mately promotes the migration of HCC cells

Article Snippet: Primary antibody against CD147, PKM2, and MCT4 were from Proteintech Group (Rosemont, IL).

Techniques: Expressing, Membrane, Migration