Journal: Gene

Article Title: Period circadian regulator 2 suppresses drug resistance to cisplatin by PI3K/AKT pathway and improves chronochemotherapeutic efficacy in cervical cancer.

doi: 10.1016/j.gene.2021.146003

Figure Lengend Snippet: Fig. 5. Overexpression of PER2 regulates clock circadian and ameliorates drug resistance through PI3K/AKT pathway in DDP-resistant cervical cancer cells. (A) Representative images of western blot. (B-M) Quantitative results of (B, C) PER2, (D, E) CLOCK, (F, G) BMAL1, (H, I) CRY1, (J, K) MRP1 and (L, M) MDR1 in Hela/ DDP as well as SiHa/DDP cells with pcDNA3.1-PER2 transfection and/or hEGF treatment. **P < 0.01; ****p < 0.0001.

Article Snippet: The sample was transferred onto the membrane at a gel volume of 1.5 mA/cm2 for 1.5 h. The membrane was added with 5% skimmed milk powder + TBST, and shaken in a shaker at room temperature lasting 1 h. The membranes were added with primary antibodies against PER2 (67513-1-Ig; Proteintech), CLOCK (18094-1-AP; Proteintech), BMAL1 (14268-1-AP; Proteintech), CRY1 (13474-1-AP; Proteintech), MRP1 (67228–1-Ig; Proteintech), MDR1 (22336-1-AP; Proteintech), PI3K (20584-1-AP; Proteintech), p-PI3K (17366S; CST), AKT (10176-2-AP; Proteintech), p-AKT (66444-1-Ig; Proteintech), Snail1 (13099-1-AP; Proteintech), Twist (25465–1-AP; Proteintech), E-cadherin (20874-1- AP; Proteintech), Vimentin (10366-1-AP; Proteintech) and GAPDH (60004-1-Ig; Proteintech) which was diluted at 1:1000 with 5% BSA + TBS, and shaken overnight at 4 ◦C.

Techniques: Over Expression, Western Blot, Transfection

Journal: Gene

Article Title: Period circadian regulator 2 suppresses drug resistance to cisplatin by PI3K/AKT pathway and improves chronochemotherapeutic efficacy in cervical cancer.

doi: 10.1016/j.gene.2021.146003

Figure Lengend Snippet: Fig. 9. Cisplatin therapy at the peak of PER2 expression ameliorates chemotherapy resistance and EMT in cervical cancer. (A) Representative images of western blot. Assessment of the expression of (B) PER2, (C) CLOCK, (D) BMAL1, (E) CRY1, (F) MRY1, (G) MRP1, (H) PI3K, (I) p-PI3K, (J) p-PI3K/PI3K, (K) AKT, (L) p-AKT, (M) p- AKT/AKT, (N) Snail, (O) Twist, (P) Vimentin and (Q) E-cadherin in tumor tissues from Hela/DDP cells-induced nude mice treated with Cisplatin at the peak or trough of PER2 expression. *P < 0.05; **p < 0.01; ***p < 0.001.

Article Snippet: The sample was transferred onto the membrane at a gel volume of 1.5 mA/cm2 for 1.5 h. The membrane was added with 5% skimmed milk powder + TBST, and shaken in a shaker at room temperature lasting 1 h. The membranes were added with primary antibodies against PER2 (67513-1-Ig; Proteintech), CLOCK (18094-1-AP; Proteintech), BMAL1 (14268-1-AP; Proteintech), CRY1 (13474-1-AP; Proteintech), MRP1 (67228–1-Ig; Proteintech), MDR1 (22336-1-AP; Proteintech), PI3K (20584-1-AP; Proteintech), p-PI3K (17366S; CST), AKT (10176-2-AP; Proteintech), p-AKT (66444-1-Ig; Proteintech), Snail1 (13099-1-AP; Proteintech), Twist (25465–1-AP; Proteintech), E-cadherin (20874-1- AP; Proteintech), Vimentin (10366-1-AP; Proteintech) and GAPDH (60004-1-Ig; Proteintech) which was diluted at 1:1000 with 5% BSA + TBS, and shaken overnight at 4 ◦C.

Techniques: Expressing, Western Blot

Journal: Annals of Translational Medicine

Article Title: Curcuma zedoaria petroleum ether extract reverses the resistance of triple-negative breast cancer to docetaxel via pregnane X receptor

doi: 10.21037/atm-21-4199

Figure Lengend Snippet: Primer sequence of the genes for qRT-PCR analysis (5'–3')

Article Snippet: The primary antibodies included the following: PXR antibody (1:50, 15607-1-AP, Proteintech, USA), MDR1 antibody (1:50, AF5185, Affinity, USA), BCRP antibody (1:50, AF5177, Affinity, USA), and CYP3A4 antibody (1:50, 18227-1-AP, Proteintech, USA).

Techniques: Sequencing

Journal: Annals of Translational Medicine

Article Title: Curcuma zedoaria petroleum ether extract reverses the resistance of triple-negative breast cancer to docetaxel via pregnane X receptor

doi: 10.21037/atm-21-4199

Figure Lengend Snippet: Effect of PECZ on the mRNA expression of drug resistance genes. The mRNA expressions of PXR (A), MDR1 (B), BCRP (C), and CYP3A4 (D) in MDA-MB-231/docetaxel cells were detected by qRT-PCR. (x¯±s, n=3), *, P<0.05; **, P<0.01 vs. control group. PECZ, petroleum ether extracts of Curcuma zedoaria; PXR, pregnane X receptor; MDR1, multidrug resistance 1; BCRP, breast cancer resistance protein; CYP3A4, cytochrome P-450.

Article Snippet: The primary antibodies included the following: PXR antibody (1:50, 15607-1-AP, Proteintech, USA), MDR1 antibody (1:50, AF5185, Affinity, USA), BCRP antibody (1:50, AF5177, Affinity, USA), and CYP3A4 antibody (1:50, 18227-1-AP, Proteintech, USA).

Techniques: Expressing, Quantitative RT-PCR

Journal: Annals of Translational Medicine

Article Title: Curcuma zedoaria petroleum ether extract reverses the resistance of triple-negative breast cancer to docetaxel via pregnane X receptor

doi: 10.21037/atm-21-4199

Figure Lengend Snippet: Effect of PECZ on the protein expression of drug resistance genes in MDA-MB-231/docetaxel cells. Representative band of each protein (A). Relative protein expression of PXR (B), MDR1 (C), BCRP (D), and CYP3A4 (E). (x¯±s, n=3), *, P<0.05; **, P<0.01 vs. control group. PECZ, petroleum ether extracts of Curcuma zedoaria; PXR, pregnane X receptor; MDR1, multidrug resistance 1; BCRP, breast cancer resistance protein; CYP3A4, cytochrome P-450.

Article Snippet: The primary antibodies included the following: PXR antibody (1:50, 15607-1-AP, Proteintech, USA), MDR1 antibody (1:50, AF5185, Affinity, USA), BCRP antibody (1:50, AF5177, Affinity, USA), and CYP3A4 antibody (1:50, 18227-1-AP, Proteintech, USA).

Techniques: Expressing

Journal: Annals of Translational Medicine

Article Title: Curcuma zedoaria petroleum ether extract reverses the resistance of triple-negative breast cancer to docetaxel via pregnane X receptor

doi: 10.21037/atm-21-4199

Figure Lengend Snippet: PECZ and docetaxel decreased the expression of drug resistance genes. Representative images of immunohistochemical showing PXR, MDR1, CYP3A4, and BCRP in tumor tissues in the different groups. Original magnification ×200, ×400; (x¯±s, n=3). PECZ, petroleum ether extracts of Curcuma zedoaria; PXR, pregnane X receptor; MDR1, multidrug resistance 1; BCRP, breast cancer resistance protein; CYP3A4, cytochrome P-450.

Article Snippet: The primary antibodies included the following: PXR antibody (1:50, 15607-1-AP, Proteintech, USA), MDR1 antibody (1:50, AF5185, Affinity, USA), BCRP antibody (1:50, AF5177, Affinity, USA), and CYP3A4 antibody (1:50, 18227-1-AP, Proteintech, USA).

Techniques: Expressing, Immunohistochemical staining

Journal: Annals of Translational Medicine

Article Title: Curcuma zedoaria petroleum ether extract reverses the resistance of triple-negative breast cancer to docetaxel via pregnane X receptor

doi: 10.21037/atm-21-4199

Figure Lengend Snippet: PECZ reduced the mRNA and protein expressions of drug resistance genes in tumor tissues. Effect of PECZ and docetaxel on the mRNA expressions of PXR (A), MDR1 (B), BCRP (C), and CYP3A4 (D) in tumor tissues. Representative band of each protein (E). Effect of PECZ and docetaxel on the protein expressions of PXR (F), MDR1 (G), BCRP (H), and CYP3A4 (I) in tumor tissues. (x¯±s, n=3); *, P<0.05; **, P<0.01 vs. model group. PECZ, petroleum ether extracts of Curcuma zedoaria; PXR, pregnane X receptor; MDR1, multidrug resistance 1; BCRP, breast cancer resistance protein; CYP3A4, cytochrome P-450.

Article Snippet: The primary antibodies included the following: PXR antibody (1:50, 15607-1-AP, Proteintech, USA), MDR1 antibody (1:50, AF5185, Affinity, USA), BCRP antibody (1:50, AF5177, Affinity, USA), and CYP3A4 antibody (1:50, 18227-1-AP, Proteintech, USA).

Techniques:

Journal: BMC Cancer

Article Title: p -Glycoprotein ABCB5 and YB-1 expression plays a role in increased heterogeneity of breast cancer cells: correlations with cell fusion and doxorubicin resistance

doi: 10.1186/1471-2407-10-388

Figure Lengend Snippet: Differential expression patterns of YB-1, c-Kit, MAPT and GST in time course . MCF-7 cells that incubated with 20 nM doxorubicin for the indicated periods of time revealed different kinetic patterns for YB-1 and GST expressions between sensitive and resistant MCF-7 cells. A: Dot blot array analysis on doxorubicin sensitive MCF-7 cells. B: Dot blot array analysis on the doxorubicin resistant MCF-7 cells. A and B: The scale on x-axis is not in proportion with time. In the time course study, actin was employed as a control for normalization, because GAPDH was regulated in doxorubicin resistant MCF-7 cells. C: Effect of doxorubicin on the expression of drug resistance related target proteins YB-1, c-Kit, ERK1/2, ERK3, FAS, MAPT, MDR1, ABCB5 and PARP-1 in the four subtypes of MCF-7 cells. 1, 2 are MCF-7 and MCF-7/vector-YB-1 respectively without treatment of doxorubicin. 3, MCF-7/vector-YB-1 with treatment of doxorubicin for 6 h (not fused cells); 4, MCF-7/vector-YB-1 with treatment of doxorubicin for 6 h (fused cells, FACS sorted R2); 5, doxorubicin resistant MCF-7 cell line. Numbers indicate a relative level of protein expression based on the level of intensity of β-actin after normalization.

Article Snippet: The protein extracts were electrophoresed by 12% SDS-PAGE, then electrically transferred onto nitrocellulose filters and probed with the following primary antibodies (specific to the proteins that may associated with acquired drug resistance): YB-1 (dilution factor 1:200, Cell Signalling), c-Kit (1:1000, Cell Signalling); ERK1/2 (1: 1000, Cell Signalling); ERK3 (1: 500, Novus Biologicals); FAS (1: 500, Santa Cruz Biotechnology); MAPT (1: 1500, Proteintech Group); and MDR1 (1: 500, Santa Cruz Biotechnology), ABCB5 (1: 2000, ProSci); PARP-1 (1: 2000, Trevigen); β-Actin (1:5000, Sigma) was used as a loading control.

Techniques: Quantitative Proteomics, Incubation, Dot Blot, Control, Expressing, Plasmid Preparation