Carfilzomib Inhibits Constitutive NF-κB Activation in Mantle Cell Lymphoma B Cells and Leads to the Induction of Apoptosis.

Mantle cell lymphoma (MCL) remains incurable and new treatments are needed, especially in the relapsed/refractory setting. We therefore investigated the effects of carfilzomib, a novel, long-acting, second-generation proteasome inhibitor, in MCL cells. Eight established MCL cell lines and freshly isolated primary MCL cells were treated with carfilzomib. Cell proliferation was assessed by a 3H-thymidine incorporation assay. Cell apoptosis was evaluated by flow cytometry with annexin V and propidium iodide. Electrophoresis mobility shift (EMSA), Western blot, and luciferase assays were used to analyze NF-κB activation and related signaling proteins. Carfilzomib inhibited growth and induced apoptosis in both established MCL cell lines and freshly isolated primary MCL cells in a dose-dependent manner. In contrast, carfilzomib was less toxic to normal peripheral blood mononuclear cells from healthy individuals. The carfilzomib-induced apoptosis of MCL cells occurred in a caspase-dependent manner through both intrinsic and extrinsic caspase pathways. In addition, carfilzomib inhibited constitutive activation of the NF-κB signaling cascade, both in MCL cell lines and primary MCL cells, by completely blocking the phosphorylation of IκBα. Our results demonstrate that carfilzomib can induce growth arrest and apoptosis in MCL cells and that the mechanism may involve the NF-κB signaling pathway.

3,4,7,8-Tetra-methyl-1,10-phenanthrolin-1-ium nitrate monohydrate.

In the crystal of the title compound, C(16)H(17)N(2) (+)·NO(3) (-)·H(2)O, the tetra-methyl-1,10-phenanthrolinium cations, nitrate anions and lattice water mol-ecules are all located on a mirror plane with the methyl H atoms of the cation equally disordered over two sites about the mirror plane. The cation, anion and water mol-ecule are linked by O-H⋯O and N-H⋯O hydrogen bonds into a sheet parallel to the bc plane. π-π stacking between phenanthroline ring systems is observed in the crystal structure, the centroid-centroid distance being 3.4745 (6) Å.

Potential effects of CRM1 inhibition in mantle cell lymphoma.

Mantle cell lymphoma (MCL) is an aggressive histotype of B-cell non-Hodgkin lymphoma. The disease has no known cure, which prompts the urgent need for novel therapeutic agents. Chromosomal region maintenance 1 (CRM1) may play a role in human neoplasia and serve as a novel target of cancer treatment. This study summarizes MCL pathogenesis and determines the involvement of CRM1 in the regulation of several vital signaling pathways contributing to MCL pathogenesis, including the pathways of cell cycle progression, DNA damage response, phosphoinositide kinase-3, nuclear factor-κB activation, and chromosomal stability. A preclinical study is also presented to compare the CRM1 status in MCL cell lines and primary MCL cells with normal B cells, as well as the therapeutic efficiency of CRM1 inhibition in MCL in vitro and in vivo, which make these agents potential targets of novel MCL treatments.

Notch1 signaling inhibits growth of EC109 esophageal carcinoma cells through downmodulation of HPV18 E6/E7 gene expression.

AIM: To investigate the role of the Notch1 signaling pathway in growth arrest of an esophageal carcinoma cell line (EC109) in vitro and the mechanism involved. METHODS: An intracellular domain of Notch1 (ICN) was transfected into cultured EC109 cells by lipofectamine transfection. Subsequently, the proliferation of the transfected cells was measured by an MTT assay. Cell cycle distribution was analyzed by flow cytometry. Human papillomavirus type 18 (HPV18) E6/E7 mRNA expression was detected by RT-PCR, and p53 protein expression was detected by Western blot. RESULTS: Activation of Notch1 signaling resulted in inhibition of EC109 cell proliferation with the induction of G(2)/M arrest, downmodulation of HPV18 E6/E7 gene expression, and upregulation of p53 expression. CONCLUSION: Repression of HPV18 E6/E7 expression by Notch1 signaling results in the activation of p53-mediated pathways with concomitant growth suppression of HPV18-positive EC109 cells.

Small molecule-driven directional movement enabling pin-hole free perovskite film via fast solution engineering.

Organolead trihalide perovskite materials have been widely used as light absorbers in efficient photovoltaic cells. Solution engineering is a fast and effective method to fabricate perovskite films. Here, we report a fast precipitation of a pin-hole free perovskite film by small molecule-driven directed diffusion engineering. Solvent molecules diffuse easily and quickly by colliding with small molecules, e.g. helium. Fully compact perovskite films and highly efficient perovskite solar cells are achieved, and the devices show remarkable stability of ca. 90% original efficiency after more than 1000 hours of testing. The small molecule driving directed diffusion offers a promising fast precipitation of a perovskite film and highly efficient, stable perovskite solar cells.

[Mechanism of radiation induced premature senescence of bone marrow stromal cells: experiment with murine bone marrow stromal cells].

OBJECTIVE: To investigate the mechanism of radiation induced premature senescence of bone marrow stromal cells induced by gamma-radiation. METHODS: Bone marrow stromal cells were isolated from the bones of mice, cultured, and divided into 2 groups: control group and irradiation group. The cells of the irradiation group were exposed to 60Co gamma-rays of the dose of 8.0 Gy. 24 h, 72 h, 1 week, and 2 weeks after irradiation MTT method was used to test the proliferation of the cells, Giemsa staining was used to observe the morphology, the percentage of cells positive in beta-galactosidase (SA-beta-Gal), a senescence associated biomarker, was detected by cytochemistry, and RT-PCR was used to detect the mRNA expression of p21(Cip1/Waf1) and p53 gene, both associated with senescence. One week after the irradiation flow cytometry was used to analyze the cell cycle distribution. RESULTS: Characteristics of mature senescence were seen in the cells of the irradiation group. One week after the irradiation the percentage of G0/G1 of the irradiation group was 89.83% +/- 0.05%, significantly higher than that of the control group (66.95% +/- 0.36%, P < 0.01). 24 h, 72 h, 1 week, and 2 weeks after irradiation the percentages of SA-beta-gal positive cells in the irradiation group were 20.33% +/- 0.03%, 34.33% +/- 0.03%, 86.33% +/- 0.02%, and 95.67% +/- 0.02% respectively, significantly higher than those of the control group (2.33% +/- 0.006%, 14.33% +/- 0.02%, 24.67% +/- 0.02%, and 45.00% +/- 0.02% respectively, all P < 0.01). The mRNA expression of p53 and p21(Cip1/Waf1) mRNA expressions in the irradiation group increased 24 h after the irradiation and lasted for two weeks. CONCLUSION: Gamma-radiation induces changes of premature senescence in bone marrow stromal cells in which p53-p21(Cip1/Waf1) manner pathways may play an important role.

[The experimental research about the effect of Prunella vulgaris L. on Raji cells growth and expression of apoptosis related protein].

OBJECTIVE: To investigate the anti-lymphoma effect of Prunella vulgaris L. in order to offer exprimental data for the treatment of lymphoma with Prunella vulgaris L. in clinic. METHODS: Effect of Prunella vulgaris L. injection on inhibition ratio of cell growth of Raji cells and IC50 were tested by MTT assay. The growth curve line of Raji cells was drawn also by MTT assay. The cellular morphology was observed by invert microscope, Giemas staining and MTT assay. The expression of apoptosis related protein bcl-2, bax was measured by immunocytochemistry and the quantitative analysis was made with figure analysis system. RESULTS: 1. Prunella vulgaris L. could obviously suppress the cell proliferation of Raji cells in a concentration-dependent manner (r = 0.97). The IC50 was 0.118 mg/ml. 2. After Raji cells was reacted with injection of Prunella vulgaris L. (50 mg/ml) , the morphlogical of apoptosis were observed by invert microscope, Giemsa staining and MTT assay. RESULTS: The results of immunocytochemistry showed that after Raji cells were treated by the injection of Prunella vulgaris L. (50 mg/ml) for 48 hours, the expression of bcl-1 was up-regulated, and the expression of bax was down-regulated. The differences between process group and control group were significant (P < 0.01). CONCLUSION: Prunella vulgaris L. can suppress the proliferation of Raji cells and may be a new anti-lymphoma drug. Inducing the apoptosis of Raji cells maybe one of anti-lymphoma mechanisms.

[Expression of pituitary tumor transforming gene in patients with acute lymphoblastic leukemia].

The aim of this study was to explore the expression of pituitary tumor-transforming gene (PTTG) in acute lymphoblastic leukemia (ALL) and its relationship with the pathogenesis of ALL, as well as study the difference of the PTTG expression in ALL patients with Ph1 chromosome and without Ph1 chromosome. The mRNA expressions of PTTG in bone marrow from 28 patients with ALL and 28 normal controls were quantitatively detected by real-time quantitative polymerase chain reaction (real-time PCR). The results indicated that the expression of PTTG mRNA was significantly higher in ALL patients (1.9428E5 ± 1.8372E5) than that in normal controls (4.5766E3 ± 1.1817E3) (P < 0.05). The expression of PTTG mRNA was higher in Ph1 chromosome positive patients. The initial expression of PTTG mRNA was lower in patients achieved complete remission than that in patients with non-complete remission. It is concluded that the overexpression of PTTG may be related to the progression and genesis of ALL. Overexpression of PTTG may be intimately related to the progression and genesis of Ph1 chromosome positive ALL. It provides a new ideas to research the pathogenesis and genic target treatment of ALL.

[Significance of platelet parameters and lactate dehydrogenase level in differential diagnosis for thrombocytosis].

This study was purposed to explore the clinical application of mean platelet volume (MPV), platelet distribution width (PDW), platelet-large cell ratio (P-LCR), lactate dehydrogenase (LDH) level in the differential diagnosis of thrombocytosis. The clinical applications of 3 platelet routine laboratory parameters (MPV, PDW, P-LCR) and LDH were examined in 1048 patients with thrombocytosis-related diseases: reactive thrombocytosis (RT), chronic myeloproliferative disease (CMPD) including chronic myeloid leukemia (CML), essential thrombocythemia (ET) and polycythemia vera (PV). Receiver operating characteristic (ROC) curve was used to predict the cause of thrombocytosis. The results indicated that there were significant differences in MCV, PDW, P-LCR and LDH level between RT and CMPD groups (p < 0.05). The area under ROC curve of PDW and P-LCR for prediction of CMPD were 0.96 (95% CI: 0.93 - 0.99) and 0.89 (95% CI: 0.84 - 0.95) respectively, and whose optimal cut-off value was 11.95%and 23.05% respectively. Three types of CMPD were characterized as follows: high P-LCR and high LDH level in chronic myeloid leukemia, whose optimal cut-off value was 424 U/L and 26.10% respectively; slightly high LDH level and high Plt count in ET, the optimal cut-off value of Plt was 939 x 109/L. In conclusion, these characteristics of MPV, PDW, P-LCR and LDH levels may be useful for simple and primary differential diagnosis of the thrombocytosis-related disease mentioned above.

[Relation of notch pathway to senescence of murine bone marrow stromal cells].

This study was purposed to investigate the relation of the Notch signaling pathway to senescence of murine bone marrow stromal cells in vitro. Intracellular domain of Notch 1 (ICN) was transfected into cultured murine bone marrow stromal cells by lipofectamine transfection. After transfection for three days the proliferation of transfected cells was measured by MTT, cell cycle distribution was analyzed by flow cytometry. The percentage of senescence associated beta-galactosidase (SA-beta-Gal) positive cells were measured by cytochemical method, and the expression rates of P53 and p21Cip1/Waf1 at gene and protein levels were analyzed by RT-PCR and Western blot respectively. The results showed that after transfection for 3 days the proliferation of murine bone marrow stromal cells was inhibited with induction of G1 arrest, the percentage of SA-beta-gal positive cells increased and the p53 and p21Cip1/Waf1 mRNA and protein expression levels were upregulated. It is concluded that the activated Notch signaling can induce premature senescence of bone marrow stromal cells through the p53-p21Cip1/Waf1 pathway.

[Expression of pituitary tumor transforming gene in patients with AML].

The aim of study was to explore the expression of pituitary tumor-transforming gene (pttg) in acute myeloid leukemia (AML) and its relationship with the pathogenesis of AML, simultaneously to investigate the difference of the pttg expression among AML different subtypes. The expressions of pttg mRNA were quantitatively detected by real-time fluorescence quantitative polymerase chain reaction (real-time PCR) in bone marrow from 47 patients with AML and 28 normal controls. The results indicated that the expression of pttg mRNA was significantly higher in AML patients [(1.1323 ± 1.3934) × 10(5)] than that in normal controls [(4.5766 ± 1.1817) × 10(3)] (p < 0.05). The expression of pttg mRNA was higher in M(3) patients than that in other AML subtypes, such as M(1), M(2), M(4), M(5). It is concluded that the overexpression of pttg may be related to the pathogenesis and progression of AML, in which the overexpression of pttg may be more intimately related to the pathogenesis and progression of M(3). This study provides a new idea to research the pathogenesis and targeted gene therapy of AML.