Correlation of CerbB-2 to Ki-67 and P53 expressions in hormone-independent breast cancer / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the relation of CerbB-2 with Ki-67 and P53 expressions in hormone-independent breast cancer.</p><p><b>METHODS</b>A total of 180 patients with breast cancer were examined for the positivity to estrogen receptor (ER) and progesterone receptor (PR), and subsequently divided into hormone-dependent group (positive for both ER and PR) and hormone-independent group (negative for both). CerbB-2, Ki-67 and P53 expressions were detected in these patients using an improved Envision immunohistochemical staining with the corresponding antibodies. Twenty-nine patients without definite immunohistochemical results of C-erbB-2 were excluded from further analysis. The 51 hormone-independent and 100 hormone-dependent cases were analyzed for the correlation of CerbB-2 to Ki-67 and P53 expressions.</p><p><b>RESULTS</b>In hormone-independent cases negative for CerbB-2, Ki-67 and P53 expressions were significantly higher than that in hormone-dependent CerbB-2-negative cases (P<0.05), but in CerbB-2-positive cases, their expressions showed no significant differences (P>0.05) with respect to hormonal dependence.</p><p><b>CONCLUSION</b>In hormone-independent CerbB-2-negative cases, the tumor cells show a higher proliferative activity and P53 expression than those in hormone-dependent cases, which can be an important reason for the high malignancy and poor prognosis of hormone-independent breast cancer negative for CerbB-2.</p>

Construction, expression and characterization of recombinant fusion protein HSA-PTH (1-34) in Pichia pastoris / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To obtain recombinant fusion protein HSA (human serum albumin)-PTH(1-34) in Pichia pastoris.</p><p><b>METHODS</b>HSA and PTH(1-34) cDNA were obtained with PCR and the DNA segments were cloned into vector pPIC9 with linker. The linearized plasmids were transformed GS115 competent cells treated with LiCl, and mut+ transformants were screened on MD plate. With AOX promoter and alpha-MF signal sequences leading, fusion protein was expressed in GS115. PCR and SDS-PAGE were employed to confirm the integration and expression of HSA-PTH(1-34). The fusion protein was identified by Western blotting and classical adenylate cyclase assay.</p><p><b>RESULT</b>The PCR results showed that the gene of HSA-PTH(1-34) was integrated into GS115 genome. Western bolt approved the existence of two domains of HSA and PTH(1-34). The bioactivity assay in rabbit cortical membranes indicated that HSA-PTH (1-34) activated adenylate cyclase, but the activity was lower than that of the synthetic PTH(1-34).</p><p><b>CONCLUSION</b>Active fusion protein HSA-PTH (1-34) is successfully expressed in Pichia pastoris.</p>

Inhibition of expression of vascular endothelial growth factor and its receptors in pulmonary adenocarcinoma cell by TNP-470 in combination with gemcitabine / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Angiogenesis is required for solid tumor growth and facilitates tumor progression and metastasis. The inhibition effects of O-(chloroacetyl-carbamoyl) fumagillol (TNP-470), an angiogenesis inhibitor, and gemcitabine, a chemotherapeutic agent, on expression of growth factors were investigated using human pulmonary adenocarcinoma cell line, A549. The A549 cells were divided into four groups: control group, 10(-6) mg/ml gemcitabine treated group, 10(-4) mg/ml TNP-470 treated group and gemcitabine+TNP-470 treated group. The mRNA and protein expression of vascular endothelial growth factor (VEGF) and its receptors, FMS-like tyrosine kinase-1 (FLT-1) and kinase insert domain-containing receptor (KDR), in different groups were measured. The growth of A549 cell cultured with gemcitabine or TNP-470 was inhibited in an almost dose-dependent manner. Although gemcitabine (10(-6) mg/ml) alone and TNP-470 (10(-4) mg/ml) alone had no effect on the mRNA and protein expression of VEGF and its receptors (FLT-1, KDR) in A549 cells compared to the control (P>0.05), 10(-6) mg/ml gemcitabine in combination with 10(-4) mg/ml TNP-470 had significant effect (P<0.01). Moreover, combination of the two drugs significantly inhibited the mRNA expression of VEGF, FLT-1 and KDR compared to either drug alone (P<0.05). This study suggests that combined treatment with TNP-470 plus gemcitabine may augment the antiangiogenic and antineoplastic effects in lung cancer cells in vitro.