Hydroxysafflor Yellow A Promotes HaCaT Cell Proliferation and Migration by Regulating HBEGF/EGFR and PI3K/AKT Pathways and Circ_0084443 / 中国结合医学杂志

OBJECTIVE@#To investigate the effect and possible mechanism of hydroxysafflor yellow A (HSYA) on human immortalized keratinocyte cell proliferation and migration.@*METHODS@#HaCaT cells were treated with HSYA. Cell proliferation was detected by the cell counting kit-8 assay, and cell migration was measured using wound healing assay and Transwell migration assay. The mRNA and protein expression levels of heparin-binding epidermal growth factor (EGF)-like growth factor (HBEGF), EGF receptor (EGFR), phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT), mammalian target of rapamycin (mTOR), and hypoxia-inducible factor-1α (HIF-1α) were detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot, respectively. Circ_0084443-overexpressing HaCaT cells and empty plasmid HaCaT cells were constructed using the lentiviral stable transfection and treated with HSYA. The expression of circ_0084443 was detected by qRT-PCR.@*RESULTS@#HSYA (800 µmol/L) significantly promoted HaCaT cell proliferation and migration (P<0.05 or P<0.01). It also increased the mRNA and protein expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and increased the phosphorylation levels of PI3K and AKT (P<0.05 or P<0.01). Furthermore, HSYA promoted HaCaT cell proliferation and migration via the HBEGF/EGFR and PI3K/AKT/mTOR signaling pathways (P<0.01). Circ_0084443 attenuated the mRNA expression levels of HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α (P<0.05). HSYA inhibited the circ_0084443 expression, further antagonized the inhibition of circ_0084443 on HBEGF, EGFR, PI3K, AKT, mTOR and HIF-1α, and promoted the proliferation of circ_0084443-overexpressing HaCaT cells (P<0.05 or P<0.01). However, HSYA could not influence the inhibitory effect of circ_0084443 on HaCaT cell migration (P>0.05).@*CONCLUSION@#HSYA played an accelerative role in HaCaT cell proliferation and migration, which may be attributable to activating HBEGF/EGFR and PI3K/AKT signaling pathways, and had a particular inhibitory effect on the keratinocyte negative regulator circ_0084443.

HBEGF up-regulate expression level of PD-L1 in glioblastoma cells partly through LIF / 中国药理学通报

Aim To investigate the mechanism of heparin-binding epidermal growth factor (HBEGF) in upregulating the expression of programmed death ligand 1 (PD-L1) in glioma cells by regulating leukemia inhibitory factor (LIF). Methods Pearson analysis was used to analyze the correlation between HBEGF, LIF and PD-L1 mRNA in primary gliomas, U251 and U87 cells were treated with HBEGF neutralizing antibody and HBEGF was added after HBEGF knockout, the mRNA and protein levels of LIF and PD-L1 were detected by real-time fluorescence quantitative PCR (RT-qPCR) and Western blot, HBEGF knockout cells were added with LIF neutralizing antibody on the basis of adding HBEGF, and the levels of PD-L1 mRNA and protein were detected. Results Pearson analysis showed that HBEGF, LIF, and PD-L1 mRNA were positively correlated (P < 0.05). After adding HBEGF neutralizing antibody to U251 and U87 cells, the mRNA and protein levels of LIF and PD-L1 gradually decreased with cell passage (P < 0.05). After HBEGF knockout, the mRNA and protein levels of LIF and PD-L1 decreased (P < 0.05), and the mRNA and protein levels of LIF and PD-L1 increased after HBEGF was added (P < 0.05). In HBEGF knockout cell lines, the levels of PD-L1 mRNA and protein were up-regulated after HBEGF was added (P<0.05), and the levels of PD-L1 mRNA and protein was down-regulated by LIF neutralizing antibody (P < 0.05). Conclusion The partial induction of LIF by HBEGF in glioblastoma is an intermediate process for HBEGF to maintain PD-L1 expression.

Relationship Between Serum mRNA Level of Heparin Binding Epidermal Growth Factor and Acute Coronary Syndrome Occurrence / 中国循环杂志

Objective: To evaluate the relationship between serum mRNA level of heparin binding epidermal growth factor (HB-EGF) and acute coronary syndrome (ACS) occurrence. Methods: Our research included in 2 groups: ACS group,n=50 patients and Control group,n=100 normal subjects. Serum HB-EGF mRNA level was examined by RT-PCR and the relationship between HB-EGF mRNA and ACS occurrence was assessed by Logistic regression analysis. Results: Compared with Control group, the serum HB-EGF mRNA level of ACS group was higher (0.22±0.73) vs (0.46±0.14),P<0.05. With adjusted meaningful factors of hypertension, smoking, TG, TC, LDL-C, HDL-C and BMI by single factor analysis, multi Logistic regression analysis indicated that serum HB-EGF mRNA was related to ACS occurrence (OR=5.813, 95% CI 2.342-14.426,P<0.001) which meant that upon 0.1 grey value of HB-EGF mRNA elevation, the risk of ACS occurrence may increase 4.813 folds accordingly. Conclusion: Serum HB-EGF mRNA level was related to ACS occurrence.

The Effect of Heparin Binding Epidermal Growth Factor on the Hypertrophy / Hyperplasia of Human Bladder Smooth Muscle

PUROPOSE: The bladder outlet obstruction (BOO) causes a bladder hypertrophy and/or hyperplasia with time. Heparin binding epidermal growth factor like growth factor (HB-EGF) is a potent mitogen for bladder smooth muscle cell (BMSC), causing significant cellular hypertrophy. Few studies have evaluated the effect of HB-EGF on bladder hypertrophy or hyperplasia. We studied to evaluated the effect of HB-EGF on the hypertrophy or hyperplasia of human BSMC and fibroblast. MATERIAL AND METHODS: To evaluate the change of expression of HB-EGF mRNA of human BMSC under a constant stimuli of 40cm-H20 pressure during 0, 1, 3 and 5 hours, RT-PCR was perfromed. The human BMSC and bladder fibroblasts with the addition of the recombinant HB-EGF 25ng/ml were performed 3H-thymidine and 3H-leucine incorporation assay for DNA and protein production, respectively. RESULTS: The expression of HB-EGF mRNA in BMSC under a constant stimuli of 40cm-H20 pressure was up-regulated with time (p<0.05). The optimal concentration of HB-EGF was 25ng/ml. The 3H-Lecine and 3H-thymidine activity in both human BSMC and fibroblast were significantly increased in addition of 25ng/ml of HB-EGF (p<0.05 and p<0.05, respectively). CONCLUSIONS: Constant hydrostatic pressure induces the expression of HB-EGF in human BSMC. Subsequently HB-EGF induces hypertrophy and hyperplasia of human BSMC and fibroblast in vitro. Our data support that HB-EGF is one of relevant BOO-induced growth factors.

Effects of Heparin-binding Epidermal Growth Factor on the Invasion Activity of an Oral Cancer Cell Line / Oral Science International

Heparin-binding epidermal growth factor (EGF)-like growth factor (HB-EGF), which belongs to the EGF family, has been shown to stimulate the growth of a variety of cells in an autocrine or paracrine manner. Although HB-EGF is widely expressed in tumors when compared to normal tissue, its contribution to tumor invasion is still not known. In the present study, we analyzed the effects of HB-EGF on the invasion activity of a cultured oral cancer cell line using short interfering RNA (siRNA).Oral squamous carcinoma cell lines, HSC3 and SAS, were transfected with siRNA targeting HB-EGF. Expression of HB-EGF was analyzed by real-time PCR. The invasiveness of the transfected cells was determined using a matrigel invasion assay, and MMP-9 production was measured by RT-PCR and gelatin zymography.The expression of HB-EGF was reduced in HSC3-siRNA and SAS-siRNA cells. The matrigel invasion assay demonstrated that the invasiveness of HSC3-siRNA and SAS-siRNA cells was reduced. Gelatin zymography demonstrated that in HSC3-siRNA and SAS-siRNA cells, MMP9 production was decreased.These findings suggest that HB-EGF expression is related to the invasion activity of oral cancer, particularly via regulation of MMP9.

Promoting Regeneration Effect of Heparin Binding Epidermal Growth Factor-Like Growth Factor on Rat Hepatocytes after Partial Orthotopic Liver Transplantation / 中国普外基础与临床杂志

Objective To evaluate the effect of heparin binding epidermal growth factor-like growth factor (HB-EGF)on liver regeneration after partial orthotopic liver transplantation.Methods Fourty SD rats were used to establish the model of partial orthotopic liver transplantation with ameliorated two-cuff technique.Then all the rats were divided into 2 groups:experiment group and control group.Twenty rats of experiment group were adminis- tered 500?g/kg HB-EGF via vena caudalis immediately after operation twice a day,while the same volume of saline was administered to the rats in control group.Five rats in each group were selected randomly and killed at the 6th hour,day 2,4 and 7 after operation,respectively.The serum levels of albumin(Alb)and alanine aminotransferase (ALT)in the blood sample were detected.Every liver was removed and weighed.The expression of Ki-67 was de- tected by using immunohistochemistry assay.The regeneration activity of hepatocytes was evaluated by flow cytom- etry.Results The wet weights of liver in experiment group were all significantly higher than that in control group at the 6th hour,day 2 and 4 after transplantation(P