Effect of Nucleolin on Lymphoma Proliferation by Regulating Thymidine Kinase 1 / 中国实验血液学杂志

OBJECTIVE@#To investigate the mechanism of nucleolin (NCL) involved in lymphoma proliferation by regulating thymidine kinase 1 (TK1).@*METHODS@#Twenty-three patients with diffuse large B-cell lymphoma (DLBCL) were selected and divided into initial treatment group (14 cases) and relapsed/refractory group (9 cases). Serum TK1 and C23 protein in peripheral blood mononuclear cells were detected. Cell models of CA46-NCL-KD (CA46-NCL-knockdown) and CA46-NCL-KNC (CA46-NCL-knockdown negative control) were established by lentivirus vector mediated transfection in Burkitt lymphoma cell line CA46. The half maximal inhibitory concentration (IC50) of CA46-NCL-KD, CA46-NCL-KNC, and CA46 to adriamycin were detected by cell proliferation assay (MTS). The expression of NCL mRNA and protein in CA46-NCL-KD and CA46-NCL-KNC cells were dectected by Q-PCR and Western blot, respectively. The cell cycle of CA46-NCL-KD, CA46-NCL-KNC, and CA46 cells were detected by flow cytometry. The expression of TK1 protein in CA46-NCL-KD and CA46-NCL-KNC cells was detected by an enhanced chemiluminescence (ECL) dot blot assay.@*RESULTS@#The level of serum TK1 in the initial treatment group was 0.43(0-30-1.01) pmol/L, which was lower than 10.56(2.19-14.99) pmol/L in the relapsed/refractory group (P<0-01), and the relative expression level of NCL protein in peripheral blood was also significantly lower. The IC50 of CA46-C23-KD cells to adriamycin was (0.147±0.02) μg/ml, which was significantly lower than (0.301±0.04) μg/ml of CA46-C23-KNC cells and (0.338±0.05) μg/ml of CA46 cells (P<0.05). Compared with CA46-NCL-KNC cells, the expression of NCL mRNA and protein, TK1 protein decreased in CA46-NCL-KD cells, and the proportion of S phase and G2/M phase also decreased, while G0/G1 phase increased in cell cycle.@*CONCLUSION@#The increased expression of NCL in DLBCL and CA46 cells indicates low sensitivity to drug. NCL may participate in regulation of lymphoma proliferation by affecting TK1 expression, thereby affecting the drug sensitivity.

Proteomics-based identification of cancer-associated proteins in chronic lymphocytic leukaemia

BACKGROUND: Chronic lymphocytic leukaemia (CLL) is a neoplasm of B-cells characterized by variable prognosis. Exploring the proteome of CLL cells may provide insights into the disease. Therefore, eleven proteomics experiments were conducted on eleven primary CLL samples. RESULTS: We reported a CLL proteome consisting of 919 proteins (false discovery rate (FDR) 1%) whose identification was based on the sequencing of two or more distinct peptides (FDR of peptide sequencing 1%). Mass spectrometry-based protein identification was validated for four different proteins using Western blotting and specific antibodies in different CLL samples. Small sizes of nucleolin (~57 kDa and ~68 kDa) showed a potential association with good prognosis CLL cells (n = 8, p < 0.01). Compared with normal B-cells, CLL cells over-expressed thyroid hormone receptor-associated protein 3 (THRAP3; n = 9; p = 0.00007), which is implicated in cell proliferation; and heterochromatin protein 1-binding protein 3 (HP1BP3; n = 10; p = 0.0002), which promotes cell survival and tumourogenesis. A smaller form of HP1BP3, which may correspond to HP1BP3 isoform-2, was specifically identified in normal B-cells (n = 10; p = 0.0001). HP1BP3 and THRAP3 predicted poor prognosis of CLL (p 0.05). Consistently, THRAP3 and HP1BP3 were found to be associated with cancer-related pathways (p 0.05). CONCLUSIONS: Our findings add to the known proteome of CLL and confirm the prognostic importance of two novel cancer-associated proteins in this disease.

Role of nucleolin on myocardial hypertrophy and fibrosis in type II diabetic cardiomyopathy mice / 中国病理生理杂志

AIM: To investigate the effect of nucleolin on diabetic cardiomyopathy in mice.METHODS: A type II diabetic cardiomyopathy mouse model was prepared using a cardiac-specific nucleolin-overexpressing transgenic mice.The mice were divided into wild-type mouse control group, nucleolin transgenic mouse control group, wild-type mouse diabetes group and nucleolin transgenic mouse diabetes group.Wheat germ agglutinin (WGA) fluorescent dye, Masson staining and PowerLab system detection were used to further clarify the role of nucleolin on cardiac hypertrophy, fibrosis and cardiac function in type II diabetic cardiomyopathy mice.RESULTS: Compared with wild-type mouse control group, no significant increase in blood glucose level was found, while genetical myocardial cell hypertrophy was significantly attenuated in nucleolin transgenic mouse diabetes group.The collagen fibers were also significantly reduced, and hemodynamic indexes ± dp/dtmax, left ventricular end-diastolic pressure, left ventricular systolic pressure and heart rate were also improved.The above differences were statistically significant.CONCLUSION: Nucleolin may reduce the occurrence of myocardial hypertrophy and fibrosis, thus improving the cardiac function of diabetic cardiomyopathy mice.

Effect of nucleolin on cardiac cell apoptosis in Type 2 diabetic cardiomyopathy mice / 中南大学学报(医学版)

Objective:To investigate the effect of nucleolin on cardiac cell apoptosis in Type 2 diabetic cardiomyopathy mice.Methods:Mice were fed with high-fat and high-sugar food for 20 weeks (mice were injected intraperitoneally with 60 mg/kg streptozotocin in the 5th and 6th weeks) to establish a mouse model of Type 2 diabetes.The mice were divided into 4 groups:a wild type (WT) control group,a nucleolin transgenic (TG) control group,a WT diabetic group,a TG diabetic group.Diabetesrelated indicators were detected at the end of the 8th week.At the end of the 20th week,HE staining was used to observe myocardial morphological changes;TUNEL staining and caspase-3 activity were used to detect the extent of apoptosis of cardiac myocytes.Results:The level of fasting blood glucose was significantly increased in the diabetic group than that in the control group.In WT diabetic group,myocardial disarrangement,fragmentation and dissolution were observed (determined by HE staining);cellular apoptosis (determined by TUNEL staining and caspase-3 activity) also increased markedly in the WT diabetic group.Compared with the wild mice in the diabetic group,myocardial morphological changes and cardiac myocytes apoptosis were alleviated significantly.Conclusion:Nucleolin overexpression affectes the occurrence and development of diabetic cardiomyopathy through inhibition of cardiac myocyte apoptosis.

Photodynamic effect of new photodynamic therapy combined with nucleolin silenceon cervical cancer cell line SiHa / 吉林大学学报(医学版)

Objective To study the antitumor effect of new photodynamic therapy (PDT) applying TMPyP4 combined with nucleolin silence on cervical cancer SiHa cells in vitro, and to explore an available combination treatment project for cervical cancer.Methods The SiHa Cells were divided into blank control group, RNAi group,PDT group and PDT-RNAi group.The proliferation activities of SiHa cells were assessed by Cell Counting Kit-8 (CCK8)assay.The apoptotic rates were measured by flow cytometry (FCM)with Annexin Ⅴ/PI staining. The invasiveness abilities were assessed by Transwell assay. Results Compared with blank control group, the inhibitory rates of growth of SiHa cells in RNAi group, PDT group and PDT-RNAi group were increased significantly (P<0.01);the inhibitory rates of growth of SiHa cells in PDT-RNAi group were higher than those in PDT group and RNAi group (P<0.05 ). The Q value was 1.27. Compared with blank control group, the apoptotic rates of SiHa cells in experiment groups were increased (P<0.05 ), and the late apoptotic rate in PDT-RNAi group was also increased;there were significant differences of the apoptotic rates between PDT-RNAi group and RNAi group, PDT group (P<0.05 ). Compared with blank control group, the cell invasiveness abilities of SiHa cells in experiment groups were decreased;there were significant differences of the invasiveness abilities of SiHa cells between PDT group and RNAi group (P<0.05).Conclusion New PDT shows a strong photodynamic effect on the SiHa cells,which can inhibit the proliferation and invasiveness and induce the apoptosis of SiHa cells invitro;nucleolin silence shows a good synergy effect to PDT.

Establishment and identification of myocardium-specific nucleolin trans-genic mice / 中国病理生理杂志

AIM:To construct myocardium-specific nucleolin ( Ncl) transgenic mice and to provide an animal model for the studies of the myocardial protection of nucleolin .METHODS:To create nucleolin transgenic mice , a myo-cardium-specific expression plasmid of nucleolin ( Alpha-MyHC clone 26-Ncl) was constructed .The gene type of transgenic mice was identified by PCR and the nucleolin protein level was tested by Western blotting .The myocardium morphology , heart weight index (HWI) and left ventricular pressure maximum rise rate were observed in nucleolin transgenic (TG) mice and wild-type ( WT) mice.RESULTS:We gained 4 transgenic mice (51, 52, 56 and 86 lines, only 52 line and 86 line were eugonic) by PCR.Western blotting analysis showed the expression of nucleolin up-regulated specifically in the myocardium .However , the myocardium morphology , HWI and left ventricular pressure maximum rise rate in the nucleolin transgenic mice were similar to those in the wild-type mice.CONCLUSION:We constructed myocardium-specific nucleo-lin transgenic mice successfully .

Specific expression of nucleolin in cap-stage tooth germ of mouse / 上海第二医科大学学报

Objective To investigate the specific expression of nucleolin in the development of cap-stage tooth germ of mouse embryo,and detect the possible biological function of nucleolin. Methods In situ hybridization and immunohistochemistry were performed to detect the expression of nucleolin mRNA and protein in the development of tooth germ at embryo day 15(E15).Double staining for apoptotic cells and nucleolin was employed to explore the relationship between nucleoin and apoptosis in tooth germ. Results The expression of nucleolin mRNA and protein was mainly detected in the cervical loop,inner epithelium and the underlying dental papilla.Moreover,nucleolin protein was also located in the primary enamel knot and basement membrane.It was revealed by double staining that the fluorescence images of nucleolin and the location of apoptotic cells were not overlapped. Conclusion The expression pattern of nucleolin mRNA do not completely coincide with that of nucleolin protein at the cap stage of tooth germ.The location for nucleolin protein in the basement membrane suggests that nucleolin may be involved in the reciprocal interactions between the inner epithelium and dental papilla mesenchyme,subsequently affect the morphogenesis of tooth germ.