[Regulation of Ruxolitinib on matrix metalloproteinase in JAK2V617F positive myeloroliferative neoplasms cells].

Objective: To investigate the regulation of JAK2 tyrosine kinase inhibitor ruxolitinib on extracellular matrix metalloproteinase (MMP in JAK2V617F positive myeloproliferative neoplasms (MPN) cells. Methods: ①Forty cases of newly diagnosed JAK2V617F positive MPN patients and 15 healthy volunteers as control in Baoding No.1 Hospital between January 2012 and December 2015 were enrolled in this study. JAK2V617F/JAK2 ratio was detected by real-time-PCR; the expression levels of phosphorylation protein tyrosine kinase 2 (p-JAK2) , MMP-2 and MMP-9 in pathological tissues of bone marrow were detected by immunohistochemistry. The bone marrow cells of JAK2V617F positive MPN patients were treated with ruxolitinib, then the migration ability and MMP-2, MMP-9 gene and protein expression levels were detected. ②The human erythroleukemia cell line HEL cells were treated with different concentrations of ruxolitinib (0, 50, 100, 250, 500, 1 000 nmol/L) . The cell viability was detected by CCK-8 test; cell migration ability was tested by transwell chambers. The mRNA expression levels of JAK2, MMP-2 and MMP-9 were detected by real-time-PCR. The protein expression levels of p-JAK2, MMP-2 and MMP-9 were detected by Western blot. Results: ①The expression levels of p-JAK2, MMP-2 and MMP-9 in the newly diagnosed group were significantly higher than control group respectively [ (78.56±24.55) % vs (41.59±17.29) %, P<0.05; (48.25±18.74) % vs (22.79±13.89) %, P<0.05; (53.29±19.28) % vs (15.56±14.96) %, P<0.05]. Spearman correlation analysis showed the positive correlation of MMP-2 and MMP-9 protein expression levels with JAK2V617F mutation (r=0.526, P=0.001; r=0.543, P=0.001) . ②The proliferation of HEL cells was inhibited by different concentrations of ruxolitinib in time and dose dependent manner. ③Cell migration test showed the number of cells leaked to the low chamber in MPN patients bone marrow cells and HEL cells treated with 5 nmol/L of ruxolitinib group were significantly lower than that without ruxolitinib treatment after 24 h [ (154.7±27.5) vs (320.3±67.3) , t=13.47, P<0.05; (70.7±10.5) vs (135.3±16.7) , t=13.89, P<0.05]. The mRNA and protein expression levels of JAK2, MMP-2 and MMP-9 decreased with the increased concentration of ruxolitinib. Conclusion: Ruxolitinib inhibits MPN cell migration and expression of MMP-2 and MMP-9 via JAK2 signal pathway.

Role of E2F-1 in chemosensitivity.

The E2F family of transcription factors, in partnership with DP proteins, is thought to regulate transcription of genes that encode protein products that are required for DNA synthesis, which include important cancer chemotherapeutic targets such as thymidylate synthase and dihydrofolate reductase. This study was conducted to investigate the effects of overexpression of human E2F-1 cDNA on chemosensitivity in a human fibrosarcoma cell line, HT-1080. The E2F-1-overexpressing HT-1080 cells had a shorter doubling time both in vitro and in vivo. Associated with an up-regulation of TS, E2F-1-transfected cells were more resistant to 5-fluorouracil than were untransfected cells. These E2F-1 transfectants, although resistant to fluoropyrimidines and serum deprivation, were more sensitive to etoposide, doxorubicin, and SN38 (the active metabolite of irinotecan) but not to Taxol.

Effect of phorbol 12-myristate-13-acetate on cell adhesion in SMMC-7721 human hepatocellular carcinoma cell line and its mechanism.

The gene expression of integrin a5 beta 1 and the ability of cell adhesion of SMMC-7721 hepatocellular carcinoma cell are reported to be altered in the presence of 100nM phorbol 12-myristate-13-acetate (PMA). The ability of cell adhesion to extracellular matrix-fibronectin (Fn) was 18.8%, 38.7% and 56.6% enhancement for 30.60 and 120 mins treatment with 100nM PMA, respectively, as compared with the PMA control. But after 6 and 12 hours, it decreased to 57.4% and 61.1%, whereas the adhesion to polylysine remained the same. Using sufficient amount of anti-a5 and anti-beta 1 mAb to pre-block the adhesion sites we found that cell adhesion to Fn was inhibited by approximately 40%. It seems that other integrins mediate cell adhesion to Fn besides alpha 5 beta 1. The results obtained from the Northern blot with alpha 5 cDNA probe showed that PMA could downregulate the transcription of integrin alpha 5 beta 1 gene from 10 mins to 12 hours. The transcription was lowered to 16.9% of the control after 30 mins. The inhibition rate was still 43.6% after 12-hour treatment. The possible mechanism by which PMA influences the cell adhesion and integrin gene expression is discussed here.

Immunization of neonates with trivalent oral poliomyelitis vaccine (Sabin).

A study was carried out between November 1981 and April 1982 on the immunological effect of administering trivalent live, oral polio vaccine to 200 mature healthy neonates from Henan Province, China. The initial dose of vaccine was given at 3 days of age, and 2 months thereafter antibodies to poliovirus types 1, 2, and 3, respectively, were detected in 46.7%, 60.7% and 48.6% of the neonates; after the second dose, the levels were 86.9%, 95.3%, and 97.2%, with geometric mean titres of 1:106.2, 1:349.8, and 1:232.5. Almost 100% of neonates exhibited antibodies after the fourth dose of vaccine. Eighty-two percent of the neonates excreted poliovirus for at least a week after the initial dose of vaccine, and this increased to 99% after the second dose. Seroconversion at 4 months of age was similar to that of a group of controls who received their initial dose of vaccine at 2 months of age; however, immunization of neonates induced immunity to poliovirus at the earliest possible age.