Effect and mechanism of ubiquitin-specific processing peptidase 10 on gemcitabine resistance of pancreatic cancer

• DOI: 10.3760/cma.j.issn.1673-9752.2016.06.014
• Publication Date: 2016-06-20
• Authors: Zhen Ning; Aman Wang; Jiwei Liu; Jian Du; Chi Ma; Yuanhang Wu; Lu Chang

Objective To investigate the effect and mechanism of ubiquitin-specific processing peptidase 10 (USP10) on gemcitabine resistance pancreatic cancer.

Methods (1) The 50% inhibiting concentration (IC50) USP10 in PANC-1, BxPC-3 SW1990 cell lines cancer was detected by CCK8 assay. (2) expression BxPC-3, SW1990-GEM Western blot. (3) Small interfering RNA (siRNA) SW1900 conducted, siRNA USP10siRNA were transected then respectively allocated into control group group. (4) IC50 between 2 groups (5) Colony-forming unit assay: number cloned cells counted colony-forming efficiency (CFE) calculated. (6) Cells interfered with 0.3 μg/mL for 48 hours, apoptosis flow cytometry percentage (7) cycle cytometry. (8) relative expressions proteins blot: ① P21 0, 0.30, 0.60, 1.20 72 hours detected. ② 0.30 at 24, 48, ③ P53 ④ line ⑤ (9) interaction co-immunoprecipitation. Measurement data presented as ±s. comparisons among evaluated one-way ANOVA, parirwise comparison analyzed LSD test. Comparison t test.

Results (1) lines: PANC-1 which using assay (0.070±0.040)μg, (0.120±0.010)μg (0.350±0.050)μg, a statistically significant difference them (F=10.765, P 0.05), there (t=3.765, 0.05). 2.650±0.050 1.450±0.060, respectively, showing (t=19.075, P<0.05). 3.250±0.050 1.550±0.050, (t=9.240, 0.590±0.050, 1.015±0.050, 2.050±0.050 2.850±0.050, concentration-dependence (F=34.088, 0.890±0.050, 1.225±0.030, 2.180±0.150 3.030±0.150, time dependence (F=29.650, results co-immunoprecipitation: protein complexes complexes.

Conclusion USP10 promotes primary acquired via activation P53/P21 pathway.

Key words: Pancreatic neoplasms; Gemcitabine; Resistance