Journal: Frontiers in Pharmacology

Article Title: Identification of a novel pyridine derivative with inhibitory activity against ovarian cancer progression in vivo and in vitro

doi: 10.3389/fphar.2022.1064485

Figure Lengend Snippet: Compound H42 induced the degradation of cyclin D1 via HDAC6. (A) mRNA levels of cyclin D1 at specific points were detected by qPCR. (B) mRNA levels of cyclin D1 after different concentrations of H42 treatment were detected by qPCR. (C) H42 promoted cyclin D1 degradation. (D) H42 downregulated HDAC6 expression and upregulated the acetylation of α-tubulin and HSP90 in ovarian cancer cells. (E) Protein expression was detected in HDAC6 overexpression ovarian cancer cells and shHDAC6 A2780 cells.

Article Snippet: A2780 cells were infected with lentiviruses carrying HDAC6 (shHDAC6) or empty vectors (GeneChem, Shanghai, China).

Techniques: Expressing, 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: 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: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803

doi: 10.12659/MSM.923664

Figure Lengend Snippet: Co-expression between CDCA5 and CDK1. ( A ) Functional proteins association network of CDCA5 and CDK1 and other co-expressed genes in the STRING database. ( B ) Scoring of various aspects of the interaction between CDCA5 and CDK1. ( C ) Analysis of the TCGA gastric cancer dataset by the GEPIA website to determine the correlation between CDCA5 and CDK1 expression (* P <0.05)

Article Snippet: To knockdown CDCA5 and CDK1 in GC cell line, specific small interfering RNAs (siRNA) against CDCA5 and CDK1, and scramble siRNA not targeting any annotated human genes (control) were designed and purchased from Shanghai Genechem (Genechem, Shanghai, China).

Techniques: Expressing, Functional Assay

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803

doi: 10.12659/MSM.923664

Figure Lengend Snippet: Co-expression analysis of CDCA5 and CDK1. Co-expression analysis of CDCA5 and CDK1 in clinical specimens of gastric cancer showed high correlation between the 2. ( A ) R=0.752, n=132; ( B ) R=0.715, n=90; ( C ) R=0.850, n=43; ( D ) R=0.632, n=27.

Article Snippet: To knockdown CDCA5 and CDK1 in GC cell line, specific small interfering RNAs (siRNA) against CDCA5 and CDK1, and scramble siRNA not targeting any annotated human genes (control) were designed and purchased from Shanghai Genechem (Genechem, Shanghai, China).

Techniques: Expressing

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803

doi: 10.12659/MSM.923664

Figure Lengend Snippet: Upregulation of CDCA5 and CDK1 in gastric cancer. ( A ) The gene expression profile of CDCA5 across all tumor samples and matched normal tissues, based on the expression of CDCA5 from the The Cancer Genome Atlas (TCGA) various types of cancer research cohort data sets, generated by the GEPIA website, the height of bar represents the median expression of certain tumor type or normal tissue. ( B ) The gene expression profile of CDK1 across all tumor samples and paired normal tissues. ( C ) Analysis of TCGA gastric cancer tissue data(n=408) and match TCGA normal and GTEx data (n=211) through the GEPIA website, to obtain CDCA5 expression levels. ( D ) Analysis of TCGA gastric cancer tissue data (n=408) and match TCGA normal and GTEx data (n=211) through the GEPIA website, to obtain CDK1 expression levels (Log 2 FC=1, P -value=0.01).

Article Snippet: To knockdown CDCA5 and CDK1 in GC cell line, specific small interfering RNAs (siRNA) against CDCA5 and CDK1, and scramble siRNA not targeting any annotated human genes (control) were designed and purchased from Shanghai Genechem (Genechem, Shanghai, China).

Techniques: Gene Expression, Expressing, Generated

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803

doi: 10.12659/MSM.923664

Figure Lengend Snippet: The relationship of CDCA5 and CDK1 in MGC-803 cells. The relative CDCA5 mRNA levels ( A ) and CDK1 mRNA levels ( B ) were analyzed in 3 different GC cell lines SGC-7901, BGC-823 and MGC-803 using RT-qPCR. Among which, MGC-803 cells could effectively be expressed in both CDCA5 and CDK1, and thus was chosen as the research cell line in this study. ( C ) Cells were assigned for 2 groups and transfected with the control siRNA (si-NC) and si-CDCA5, respectively. The CDCA5 mRNA level was reduced significantly compared with the control group (** P <0.01). ( D ) The CDCA5 protein level was reduced significantly compared with the control (* P <0.05). The results suggested the si-CDCA5 fragment was effective. ( E ) Cells transfected with three different CDCA5-specific siRNAs were named as si-CDK1–1, si-CDK1–2 and si-CDK1–3, respectively. Group transfected with scramble siRNA was named as si-NC and worked as a control. Compared with the si-NC control group, the mRNA level of CDK1 in the 3 siRNA interfering groups were all reduced, among which, the si-CDK1–2 group reduced most significantly (* P <0.05 and ** P <0.01). Thus, we selected si-CDK1–2 as the specific siRNA fragment against CDK1 for the following experiments and renamed si-CDK1. ( F ) The protein expression levels of CDK1 in the si-CDK1 group and the control. Consistent with the result in ( E ), the protein expression level reduced significantly in the si-CDK1 group (* P <0.05).

Article Snippet: To knockdown CDCA5 and CDK1 in GC cell line, specific small interfering RNAs (siRNA) against CDCA5 and CDK1, and scramble siRNA not targeting any annotated human genes (control) were designed and purchased from Shanghai Genechem (Genechem, Shanghai, China).

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

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803

doi: 10.12659/MSM.923664

Figure Lengend Snippet: Relationship between CDK1 expression and CDCA5 expression. To explore the relationship between CDK1 and CDCA5, cells were knocked down one gene and checked the mRNA and protein expression level changes of the other gene. ( A ) Relative mRNA expression levels of CDK1 in cells transfected with si-NC and si-CDCA5 by quantitative real-time polymerase chain reaction (RT-qPCR) (** P <0.01). ( B ) Relative protein expression levels of CDK1 in cells transfected with si-NC and si-CDCA5 by western blot (* P <0.05). ( C ) Relative mRNA expression levels of CDCA5 in cells transfected with si-NC and si-CDK1 by RT-qPCR (* P <0.05). ( D ) Relative protein expression levels of CDCA5 in cells transfected with si-NC and si-CDK1 by western blot (* P <0.05).

Article Snippet: To knockdown CDCA5 and CDK1 in GC cell line, specific small interfering RNAs (siRNA) against CDCA5 and CDK1, and scramble siRNA not targeting any annotated human genes (control) were designed and purchased from Shanghai Genechem (Genechem, Shanghai, China).

Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Western Blot

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803

doi: 10.12659/MSM.923664

Figure Lengend Snippet: CDK1 or CDCA5 knockdown reduced cell proliferation in MGC-803. ( A ) The CCK-8 measurement results for the cells transfected with si-NC, si-CDCA5 and si-CDK1 at 0, 24, 48, and 72 hours, respectively. ( B ) The cell colon formation results for cells transfected with si-NC, si-CDCA5 and si-CDK1. ( C ) The cell colony number for cells transfected with si-NC, si-CDCA5 and si-CDK1 (** P <0.01).

Article Snippet: To knockdown CDCA5 and CDK1 in GC cell line, specific small interfering RNAs (siRNA) against CDCA5 and CDK1, and scramble siRNA not targeting any annotated human genes (control) were designed and purchased from Shanghai Genechem (Genechem, Shanghai, China).

Techniques: Knockdown, CCK-8 Assay, Transfection

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803

doi: 10.12659/MSM.923664

Figure Lengend Snippet: CDK1 or CDCA5 knockdown suppressed cell migration in MGC-803. ( A ) The cell scratch assay results for the cells transfected with si-NC, si-CDCA5 and si-CDK1 at 0, 24, 48 hours, respectively. ( B ) Relative cell migration ability of cells transfected with si-NC, si-CDCA5 and si-CDK1 at 48 hours (* P <0.05 and ** P <0.01). ( C ) The Transwell migration assay for cells transfected with si-NC, si-CDCA5 and si-CDK1, respectively. ( D ) The cell number per field for cells transfected with si-NC, si-CDCA5 and si-CDK1 (* P <0.05 and ** P <0.01).

Article Snippet: To knockdown CDCA5 and CDK1 in GC cell line, specific small interfering RNAs (siRNA) against CDCA5 and CDK1, and scramble siRNA not targeting any annotated human genes (control) were designed and purchased from Shanghai Genechem (Genechem, Shanghai, China).

Techniques: Knockdown, Migration, Wound Healing Assay, Transfection, Transwell Migration Assay

Journal: Medical Science Monitor : International Medical Journal of Experimental and Clinical Research

Article Title: Cyclin-Dependent Kinase 1 (CDK1) is Co-Expressed with CDCA5: Their Functions in Gastric Cancer Cell Line MGC-803

doi: 10.12659/MSM.923664

Figure Lengend Snippet: CDK1 or CDCA5 knockdown inhibited cell invasion in MGC-803. ( A ) The Transwell invasion assay for cells transfected with si-NC, si-CDCA5 and si-CDK1, respectively. ( B ) The cell number per field for cells transfected with si-NC, si-CDCA5 and si-CDK1 (** P <0.01).

Article Snippet: To knockdown CDCA5 and CDK1 in GC cell line, specific small interfering RNAs (siRNA) against CDCA5 and CDK1, and scramble siRNA not targeting any annotated human genes (control) were designed and purchased from Shanghai Genechem (Genechem, Shanghai, China).

Techniques: Knockdown, Transwell Invasion Assay, Transfection