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1.
J Cell Mol Med ; 22(6): 3238-3245, 2018 06.
Article in English | MEDLINE | ID: mdl-29602203

ABSTRACT

Emerging evidence has validated the vital role of long non-coding RNA (lncRNA) in the chemoresistance of cancer treatment. In the present study, we investigate the function of lncRNA NR2F1-AS1 on oxaliplatin (OXA) resistance of hepatocellular carcinoma (HCC) and discover the underlying molecular mechanism. Results revealed that lncRNA NR2F1-AS1 was up-regulated in oxaliplatin-resistant HCC tissue and cells using microarray analysis and RT-PCR. Meanwhile, ABCC1 protein was overexpressed in OXA-resistant HCC cells (Huh7/OXA and HepG2/OXA). In vitro, NR2F1-AS1 knockdown reduced the invasion, migration, drug-resistant gene (MDR1, MRP5, LRP1) and IC50 value in Huh7/OXA and HepG2/OXA cells. In vivo, NR2F1-AS1 knockdown decreased the tumour weight of HCC cells. Bioinformatics tools and luciferase reporter assay confirmed miR-363 targeted the 3'-UTR of NR2F1-AS1 and ABCC1 mRNA, presenting that NR2F1-AS1 promoted ABCC1 expression through endogenous sponging miR-363. In summary, results conclude that NR2F1-AS1 regulates HCC OXA resistance through targeting miR-363-ABCC1 pathway, providing a vital theoretic mechanism and therapeutic target for HCC chemoresistance.


Subject(s)
Carcinoma, Hepatocellular/drug therapy , Liver Neoplasms/drug therapy , MicroRNAs/genetics , Multidrug Resistance-Associated Proteins/genetics , RNA, Long Noncoding/genetics , Carcinoma, Hepatocellular/genetics , Carcinoma, Hepatocellular/pathology , Cell Movement/genetics , Cell Proliferation/genetics , Drug Resistance, Neoplasm/genetics , Gene Expression Regulation, Neoplastic , Hep G2 Cells , Heterografts , Humans , Liver Neoplasms/genetics , Liver Neoplasms/pathology , Neoplasm Invasiveness/genetics , Neoplasm Invasiveness/pathology , Oxaliplatin/administration & dosage , Oxaliplatin/adverse effects
2.
China Journal of Endoscopy ; (12): 36-40, 2017.
Article in Chinese | WPRIM (Western Pacific) | ID: wpr-664346

ABSTRACT

Objective To investigate the operation process of the technology, safety and operability of total laparoscopic resection for colorectal cancer by natural orifice specimen extraction (NOSE). Methods 40 patients with colorectal cancer who met the inclusion criteria of NOSE method from April 2015 to June 2017 were randomly divided into control group (traditional laparoscope) and experimental group (NOSE group), 20 cases in each. The intraoperative and postoperative quality of life between the two groups were statistically analyzed. Results All the patients completed the target operation, and no other operative methods were transferred. No complications occurred in either group. There were statistically difference (P < 0.05) between the two groups of patients in the two indicators (time and blood loss), there was no statistically significant difference in hospital time (P > 0.05), there was statistically difference (P < 0.05) between the two groups of quality of life score (SF-36 scale) in somatic function, role function, pain, cognitive and overall health status of five dimensions, the NOSE group was superior to the traditional laparoscopic group. Conclusion There are advantages in totally laparoscopic colorectal cancer treated with whole NOSE method. The overall health is good, few restrictions on daily work and life, quicker recovery of physical function and role function. Therefore, the application can be promoted if the condition is allowed.

3.
World J Gastroenterol ; 12(27): 4401-5, 2006 Jul 21.
Article in English | MEDLINE | ID: mdl-16865786

ABSTRACT

AIM: To construct the expression vectors for prokaryotic and eukaryotic human augmenter of liver regeneration (hALR) and to study their biological activity. METHODS: hALRcDNA clone was obtained from plasmid pGEM-T-hALR, and cDNA was subcloned into the prokatyotic expression vector pGEX-4T-2. The recombinant vector and pGEX-4T-2hALR were identified by enzyme digestion and DNA sequencing and transformed into E coli JM109. The positively selected clone was induced by the expression of GST-hALR fusion protein with IPTG, then the fusion protein was purified by glutathine s-transferase (GST) sepharose 4B affinity chromatography, cleaved by thrombin and the hALR monomer was obtained and detected by measuring H thymidine incorporation. RESULTS: The product of PCR from plasmid pGEM-T-hALR was examined by 1.5% sepharose electrophoresis. The specific strap was coincident with the theoretical one. The sequence was accurate and pGEX-4T-hALP digested by enzymes was coincident with the theoretical one. The sequence was accurate and the fragment was inserted in the positive direction. The recombinant vector was transformed into E coli JM109. SDS-PAGE proved that the induced expressive fusion protein showed a single band with a molecular weight of 41 kDa. The product was purified and cleaved. The molecular weights of GST and hALR were 26 kDa, 15 kDa respectively. The recombinant fusion protein accounted for 31% of the total soluble protein of bacterial lysate. HALR added to the culture medium of adult rat hepatocytes in primary culture and HepG2 cell line could significantly enhance the rate of DNA synthesis compared to the relevant control groups (P < 0.01). CONCLUSION: Purified hALR has the ability to stimulate DNA synthesis of adult rat hepatocytes in primary culture and HepG2 cells in vitro, and can provide evidence for its clinical application.


Subject(s)
Cytochrome Reductases/genetics , Cytochrome Reductases/isolation & purification , Gene Expression Regulation, Enzymologic , Genetic Vectors/genetics , Animals , Carcinoma, Hepatocellular/pathology , Cell Proliferation/drug effects , Cells, Cultured , Cytochrome Reductases/physiology , DNA, Complementary/analysis , DNA, Complementary/genetics , DNA, Complementary/metabolism , Escherichia coli/enzymology , Glutathione Transferase/genetics , Green Fluorescent Proteins/genetics , Hepatocytes/drug effects , Humans , Liver Neoplasms/pathology , Male , Oxidoreductases Acting on Sulfur Group Donors , Rats , Rats, Wistar , Recombinant Fusion Proteins/genetics , Tumor Cells, Cultured
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