[Experimental research of neutrophil gelatinase-associated lipocalin siRNA encapsulated by urocanic acid-coupled chitosan on colon cancer cells].

Autor:	Shen Z; Department of Coloproctology, The Third Peoples' Hospital of Hangzhou, Hangzhou 310009, China., Xu K; Department of Coloproctology, The Third Peoples' Hospital of Hangzhou, Hangzhou 310009, China., Wang H; Department of Coloproctology, The Third Peoples' Hospital of Hangzhou, Hangzhou 310009, China., Yang G; Department of Coloproctology, The Third Peoples' Hospital of Hangzhou, Hangzhou 310009, China., Pan J; Analysis and Testing Center, Zhejiang Chinese Medical University, Hangzhou 310053, China., Li M; Analysis and Testing Center, Zhejiang Chinese Medical University, Hangzhou 310053, China., Qiu J; Department of Coloproctology, The Third Peoples' Hospital of Hangzhou, Hangzhou 310009, China., Wu W; Department of Coloproctology, The Third Peoples' Hospital of Hangzhou, Hangzhou 310009, China., Zhang Y; Department of Medical Imaging, Second Affiliated Hospital Zhejiang University School of Medicine, Hangzhou 310009, China., Zhang X; Department of Coloproctology, The Third Peoples' Hospital of Hangzhou, Hangzhou 310009, China. legandsky@163.com.
Jazyk:	čínština
Zdroj:	Zhonghua wei chang wai ke za zhi = Chinese journal of gastrointestinal surgery [Zhonghua Wei Chang Wai Ke Za Zhi] 2017 Jun 25; Vol. 20 (6), pp. 694-700.
Abstrakt:	Objective: To explore the impact of neutrophil gelatinase-associated lipocalin (NGAL) knockdown by NGAL siRNA encapsulated with urocanic acid-modified chitosan nanoparticles (UAC) on the proliferation, migration and apoptosis of human colon cancer cells. Methods: NGAL siRNA was encapsulated by UAC and chitosan (CTS) respectively, and then was transfected into human colon cancer cell lines HT29. The NGAL mRNA was detected by real-time quantitative PCR (RT-QPCR). Relationships of NGAL gene silencing with the proliferation, migration and apoptosis of HT29 cell were analyzed. Results: Under the fluorescence microscope, the transfection efficiency of siRNA in UAC group was (37.52±7.17)%, which was significantly higher than (11.32±3.39)% in CTS group (t=6.102, P=0.005). Forty-eight hours after transfection, RT-QPCR examination showed that the level of NGAL mRNA expression was 0.350 in UAC group and 0.529 in CTS group with significant difference (t=-3.743, P=0.02), meanwhile both levels were significantly lower as compared to control group(F=163.538, P<0.001). Proliferation analysis revealed that after silencing NGAL gene, proliferation rate of UAC group and CTS group was slightly lower than control group, and no significant differences were found (F=9.520, P=0.438). However, migration assay demonstrated that the 24-hour migration rate of UAC group and CTS group was significantly lower than that of control group (F=6.756, P=0.029), meanwhile the migration rate of UAC group was slightly lower than that of CTS group [(77.90±7.14)% vs. (87.67±3.98)%, t=-1.704, P=0.164]. Apoptosis detection revealed that the apoptosis rate in UAC group was significantly higher than that in CTS group and the control group 2 days after transfection [(15.800±1.054)% vs. (12.900±0.656)%, (11.933±1.914)%, F=7.004, P=0.027]. Conclusions: The encapsulated ability and transfection efficiency of chitosan modified by urocanic acid elevate significantly. Silencing NGAL gene by UAC carrier can down-regulate the expression of NGAL mRNA in HT29 colon cell line, inhibit their migration and facilitate their apoptosis.
Databáze:	MEDLINE
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