Journal: Cancers

Article Title: Implication of COPB2 Expression on Cutaneous Squamous Cell Carcinoma Pathogenesis

doi: 10.3390/cancers14082038

Figure Lengend Snippet: Effect of COPB2 knockdown on biologic behavior of cutaneous squamous cell carcinoma (cSCC) cells: ( A ) (i) COPB2 protein expression decreased predominantly after COPB2 knockdown in both HSC-1 and A431 cells (Magnification, 400×; Scale bar, 50 μm); (ii) Proliferation ability of both HSC-1 and A431 cells decreased significantly after COPB2 knockdown at the indicated time points (Both p < 0.001, Mann-Whitney U-test). ( B ) (i) Representative patterns of cell invasion in each group of cSCC cells (Magnification, 100×; Scale bar, 200 μm). (ii) Invasion ability of both HSC-1 and A431 cells decreased significantly after COPB2 knockdown (Both p = 0.002, Mann-Whitney U-test). ( C ) Apoptotic cells increased predominantly after COPB2 knockdown in both HSC-1 and A431 cells. ( D ) (i) Tumor nodules of xenograft mouse models obtained from control and shCOPB2 groups of A431 cells (Scale bar, 1 cm). (ii) Volume of the tumor nodules increased significantly in control group compared to that in the COPB2 knockdown group at the indicated days (* p = 0.008, Mann-WhitneyU-test). (iii) Representative patterns of apoptotic cells in tumor nodules from each group of A431 cells-xenograft models (Magnification, 400×; Scale bar, 50 μm). Apoptotic cells were predominantly higher in the COPB2 knockdown group than in the control group in A431 cells ( p < 0.001, Mann-Whitney U-test).

Article Snippet: COPB2 knockdown stable HSC-1 and A431, as well as related control cells, were established using pGFP-C-shCOPB2 lentiviral particles (OriGene, Rockville, MD, USA) and maintained in Roswell Park Memorial Institute (RPMI) 1640 medium supplemented with 10% fetal bovine serum (FBS; Invitrogen, Waltham, MA, USA) and 1% streptomycin/penicillin (Gibco, Waltham, MA, USA).

Techniques: Expressing, MANN-WHITNEY

Journal: Molecules (Basel, Switzerland)

Article Title: Standardized Pomegranate (Pomella ® ) and Red Maple (Maplifa ® ) Extracts and Their Phenolics Protect Type I Collagen by the Inhibition of Matrix Metalloproteinases, Collagenase, and Collagen Cross-Linking.

doi: 10.3390/molecules27227919

Figure Lengend Snippet: Figure 6. Protein expression levels of metalloproteinase 13 (MMP-13) on NTERT and A431 whole cell lysates without any treatment (A) and significant increase of MMP-13 expression in A431 compared to NTERT (B). Measurement of MMP13 expression on A431 upon treatment with Pomella® and Maplifa® extracts (both at 12.5 µg/mL), their pure compounds PA and GA (both at 12.5 µM) and their 1:1 combination each at 12.5 µM. The inhibitory effects were determined by the Western blot assay using A431 cell lysates (C). Data are expressed as means ± standard deviation (S.D.) from three independent experiments performed in duplicate (B,D). Statistically significant difference was considered * p < 0.05, ** p < 0.01, *** p < 0.001 when compared to the control group and ## p < 0.01, ### p < 0.001 when compared between individual treatment groups.

Article Snippet: Epidermoid carcinoma A431 cell lines were purchased from Angio-Proteomie (Boston, MA, USA) and cultured in DMEM supplemented with 5% FBS.

Techniques: Expressing, Western Blot, Standard Deviation, Control

Journal: Molecules (Basel, Switzerland)

Article Title: Standardized Pomegranate (Pomella ® ) and Red Maple (Maplifa ® ) Extracts and Their Phenolics Protect Type I Collagen by the Inhibition of Matrix Metalloproteinases, Collagenase, and Collagen Cross-Linking.

doi: 10.3390/molecules27227919

Figure Lengend Snippet: Figure 7. Induction of type I collagen (collagen 1A1) protein expression levels and modulation of matrix metalloproteinase 9 (MMP-9) after 48 h treatment of A431 with Maplifa® (25 µg/mL), Pomella® (12.5 µg/mL), Maplifa® (25 µg/mL) + Pomella (12.5 µg/mL), GA (12.5 µM), PA (12.5 µM) and GA+PA (12.5 µM) by Western blotting. The inductive or inhibitory effects were analyzed by Western blot assay of A431 cell lysates (A). Data are expressed as means ± standard deviation (S.D.) from three independent experiments each performed in duplicate (B,C for collagen 1 A1 and MMP9 respectively). Statistically significant difference was considered, not significant (ns) compared to control, * p < 0.05, ** p < 0.01, *** p < 0.001 when compared to the control group and ## p < 0.01, ### p < 0.001, #### p < 0.0001 when compared between individual treatment groups.

Article Snippet: Epidermoid carcinoma A431 cell lines were purchased from Angio-Proteomie (Boston, MA, USA) and cultured in DMEM supplemented with 5% FBS.

Techniques: Expressing, Western Blot, Standard Deviation, Control

Journal: BMC Cancer

Article Title: Aberrant methylation of the M-type phospholipase A 2 receptor gene in leukemic cells

doi: 10.1186/1471-2407-12-576

Figure Lengend Snippet: PLA2R1 gene methylations in different human cell lines

Article Snippet: In addition, genomic DNA from Raji (human B-cell leukemia), MCF7 (human mammary adenocarcinoma), and A431 (human melanoma) cell lines was purchased from BioCat GmbH (Heidelberg, Germany).

Techniques: Methylation

Journal: Cancer Management and Research

Article Title: Grape Seed Proanthocyanidins (GSPs) Inhibit the Development of Cutaneous Squamous Cell Carcinoma by Regulating the hsa_circ_0070934/miR-136-5p/PRAF2 Axis

doi: 10.2147/CMAR.S302084

Figure Lengend Snippet: GSPs regulated hsa_circ_0070934 expression and the malignant progression of CSCC. A431 and SCL-1 cells were treated with different concentrations of GSPs for 24 h. ( A and B ) The expression of hsa_circ_0070934 was measured by qRT-PCR. MTT assay ( C and D ) and colony formation assay ( E ) were used to detect cell viability and the number of colonies to assess cell proliferation. ( F – H ) Flow cytometry was performed to determine the cell cycle process and apoptotic cells. ( I and J ) The numbers of migrated and invaded cells were evaluated by transwell assay. ( K ) The protein levels of PCNA, c-caspase 3/caspase 3 and MMP-3 were examined using WB analysis. * P < 0.05.

Article Snippet: Human CSCC cell lines (A431 and SCL-1) and normal epidermal cells (HaCaT) were obtained from Biovector National Typical Culture Center (NTCC, Beijing, China).

Techniques: Expressing, Quantitative RT-PCR, MTT Assay, Colony Assay, Flow Cytometry, Transwell Assay

Journal: Cancer Management and Research

Article Title: Grape Seed Proanthocyanidins (GSPs) Inhibit the Development of Cutaneous Squamous Cell Carcinoma by Regulating the hsa_circ_0070934/miR-136-5p/PRAF2 Axis

doi: 10.2147/CMAR.S302084

Figure Lengend Snippet: GSPs inhibited CSCC cell progression by regulating hsa_circ_0070934. ( A ) The transfection efficiency of hsa_circ_0070934 overexpression vector was assessed by detecting hsa_circ_0070934 expression in A431 and SCL-1 cells using qRT-PCR. ( B – L ) A431 and SCL-1 cells were transfected with Vector or hsa_circ_0070934 overexpression vector, and then treated with GSPs. Non-transfected cells were used as GSPs group. Non-transfected and non-treated cells were used as control group. ( B ) QRT-PCR was employed to detect hsa_circ_0070934 expression. Cell viability and the number of colonies were determined using MTT assay ( C and D ) and colony formation assay ( E ) to evaluate cell proliferation. ( F – H ) Cell cycle process and apoptotic cells were measured by flow cytometry. ( I – K ) Transwell assay was performed to assess the numbers of migrated and invaded cells. ( L ) WB analysis was utilized to test the protein levels of PCNA, c-caspase 3/caspase 3 and MMP-3. * P < 0.05.

Article Snippet: Human CSCC cell lines (A431 and SCL-1) and normal epidermal cells (HaCaT) were obtained from Biovector National Typical Culture Center (NTCC, Beijing, China).

Techniques: Transfection, Over Expression, Plasmid Preparation, Expressing, Quantitative RT-PCR, Control, MTT Assay, Colony Assay, Flow Cytometry, Transwell Assay

Journal: Cancer Management and Research

Article Title: Grape Seed Proanthocyanidins (GSPs) Inhibit the Development of Cutaneous Squamous Cell Carcinoma by Regulating the hsa_circ_0070934/miR-136-5p/PRAF2 Axis

doi: 10.2147/CMAR.S302084

Figure Lengend Snippet: Hsa_circ_0070934 sponged miR-136-5p to regulate the progression of GSPs-treated CSCC cells. A431 and SCL-1 cells were transfected with Vector, hsa_circ_0070934, hsa_circ_0070934 + NC mimic or hsa_circ_0070934 + miR-136-5p mimic, and then treated with GSPs. ( A ) MiR-136-5p expression was measured by qRT-PCR. MTT assay ( B and C ) and colony formation assay ( D ) were performed to measure cell viability and the number of colonies to evaluate cell proliferation. ( E – G ) Cell cycle process and apoptotic cells were analyzed using flow cytometry. ( H – K ) The numbers of migrated and invaded cells were determined using transwell assay. ( L ) WB analysis was used to measure the protein levels of PCNA, c-caspase 3/caspase 3 and MMP-3. * P < 0.05.

Article Snippet: Human CSCC cell lines (A431 and SCL-1) and normal epidermal cells (HaCaT) were obtained from Biovector National Typical Culture Center (NTCC, Beijing, China).

Techniques: Transfection, Plasmid Preparation, Expressing, Quantitative RT-PCR, MTT Assay, Colony Assay, Flow Cytometry, Transwell Assay

Journal: Cancer Management and Research

Article Title: Grape Seed Proanthocyanidins (GSPs) Inhibit the Development of Cutaneous Squamous Cell Carcinoma by Regulating the hsa_circ_0070934/miR-136-5p/PRAF2 Axis

doi: 10.2147/CMAR.S302084

Figure Lengend Snippet: MiR-136-5p inhibited CSCC cell progression by targeting PRAF2. ( A and B ) The transfection efficiency of pcDNA PRAF2 overexpression vector was evaluated by detecting the mRNA and protein expression of PRAF2 in A431 and SCL-1 cells using qRT-PCR and WB analysis. ( C and D ) A431 and SCL-1 cells were transfected with NC mimic, miR-136-5p mimic, miR-136-5p mimic + pcDNA or miR-136-5p mimic + PRAF2. ( C and D ) The mRNA and protein expression of PRAF2 was measured by qRT-PCR and WB analysis. Cell viability and the number of colonies were analyzed using MTT assay ( E and F ) and colony formation assay ( G ) to assess cell proliferation. ( H – J ) Flow cytometry was utilized to detect cell cycle process and apoptotic cells. ( K – M ) Transwell assay was employed to evaluate the numbers of migrated and invaded cells. ( N ) The protein levels of PCNA, c-caspase 3/caspase 3 and MMP-3 were detected using WB analysis. * P < 0.05.

Article Snippet: Human CSCC cell lines (A431 and SCL-1) and normal epidermal cells (HaCaT) were obtained from Biovector National Typical Culture Center (NTCC, Beijing, China).

Techniques: Transfection, Over Expression, Plasmid Preparation, Expressing, Quantitative RT-PCR, MTT Assay, Colony Assay, Flow Cytometry, Transwell Assay

Journal: Cancer Management and Research

Article Title: Grape Seed Proanthocyanidins (GSPs) Inhibit the Development of Cutaneous Squamous Cell Carcinoma by Regulating the hsa_circ_0070934/miR-136-5p/PRAF2 Axis

doi: 10.2147/CMAR.S302084

Figure Lengend Snippet: GSPs restrained CSCC tumor growth via regulating the hsa_circ_0070934/miR-136-5p/PRAF2 axis. A431 cells were injected into nude mice, and then the mice were given a gavage of 200 mg/kg GSPs daily (0 mg/kg GSPs was used as control group) when the tumor volume reached about 100 mm 3 . ( A ) Tumor volume was measured every 4 days. ( B and C ) After the tumor was removed, the tumor was photographed and weighted. ( D and E ) The expression of hsa_circ_0070934 and miR-136-5p was measured by qRT-PCR. ( F and G ) The mRNA and protein expression of PRAF2 was determined using qRT-PCR and WB analysis. * P < 0.05.

Article Snippet: Human CSCC cell lines (A431 and SCL-1) and normal epidermal cells (HaCaT) were obtained from Biovector National Typical Culture Center (NTCC, Beijing, China).

Techniques: Injection, Control, Expressing, Quantitative RT-PCR