Effect of Homoharringtonine on Bone Morrow CD34^＋CD7^＋ Cells in Chronic Granulocytic Leukemia

Objective： To explore the effect of homoharringtonine （HHT） on bone morrow CD34^＋CD7^＋ cells in chronic granulocytic leukemia （CGL）. Methods： The changes of bone morrow CD34^＋CD7^＋ cells were observed after the treatment of HHT in 23 cases with CGL. The proliferation and apoptosis of CD34^＋CD7^＋ cells treated with HHT in vitro were studied. Results： The proportion of CD34^＋CD7^＋ cells in CGL （0.145±0.021） was higher than that of normal control （0.052±0.013）. The proportion of CD34^＋CD7^＋ cells in patients who got cytogenetic responses to HHT （0.072±0.020） decreased remarkably, but not in those patients who did not got cytogenetic responses to HHT, （0.137±0.023）. the proliferation of CD34^＋ cells was inhibited and the proportion of CD34^＋CD7^＋ cells decreased after cultured with HHT （0.134 in 24 h, 0.126 in 48 h and 0.102 in 72）. The apoptosis rate of CD34^＋CD7^＋ cells was higher than that in CD34^＋CDT cells （35.39%±4.39% versus 24.57%±4.01%, P〈0.05） 72 h after culture with HHT. Conclusion： The proportion of CD34^＋CD7^＋ cells in CGL was higher than that of normal control and HHT may inhibit the proliferation and induce apoptosis of bone marrow CD34^＋CD7^＋ cells.     

DC-CIK细胞的生物学活性及抗淋巴瘤细胞的作用

Objective: To investigate the proliferation capabilities, immunophenotype changes, level of secreted cytokines and activities against lymphoma cells under the condition that cytokine-induced killer (CIK) cells co-cultured with dendritic cells (DC) in vitro. Methods: DC and CIK cells were induced from peripheral blood mononuclear cells of healthy volunteers. They were co-cultured meanwhile CIK cells were cultured alone as controls. Increased number of cells were counted by tapan-blue staining, killing activities were detected by MTT assay, immunophenotype changes were analyzed by flow cytometry, the IL-12 and INF-γ levels of the cultured supernatants were detected by ELISA kits. Results: The proliferation capabilities of DC-CIK cells were significantly higher than that of CIK cells (P < 0.05). Under the same condition, the ratio of double positive cells such as CD3 CD8 , CD3 CD56 in CIK cells was significantly enhanced by co-cultured with DC cells (P < 0.05). The level of IL-12 and INF-γ secreted in supernatants was increased noticeably by co-cultured DC-CIK cells on day 3 compared to CIK cells which were cultured alone (P < 0.01 and P < 0.05). Within the effector-target ratio range between 5:1 to 40:1, the activi-ties against lymphoma cells of DC-CIK cells were much higher than that of CIK cells (P < 0.05), and this effect was showed a positive correlation with the effector-target ratio. Conclusion: The proliferation capabilities, the level of secreted cytokines and the activities against lymphoma cells of DC-CIK cells were significantly higher than those of CIK cells. The research might provides theoretical and experimental basis for clinical immunotherapy of DC-CIK cells.     

NK/T细胞淋巴瘤细胞凋亡和细胞增殖特征及意义

Objective:The aim was to study the features and clinical significance of cell apoptosis and proliferation of NK/T cell lymphoma.Methods:TdT-mediated dUTP nick end labeling and immunohistochemical Streptavidin-peroxidase method were used to study cell apoptosis and the expression of proliferation cell nuclear antigen in 25 NK/T cell lymphoma and 10 reactive lymphoid tissues.Results:Apoptotic index(AI) and proliferative index(PI) averaged(1.92%±0.86%) and(41.48%±5.10%) respectively in the 25 NK/T cell lymphom...     

宫颈癌中细胞增殖和细胞凋亡的关系研究

Objective To study the relationship between cell proliferation and apoptosis in cervical carcinoma and its clinical significance.Methods The cell proliferation and apoptosis of cervical epithelial cells in archival formalin-fixed,paraffin-embedded tissue sections of normal cervix ,cervical intraepithelial neoplasms(CN) and cervical squamous carcinoma were tested by using immunohistochemistry assay and DNA nick end-labeling technigue.The proliferation index(PI) and apoptosis index(AI) were calculated and their correlation with clinical and pathological data was analyzed. Results PI was gradually increased,but the AI and AI/PI ratio decreased from normal cervical epithelium,CIN to cervical carcinoma. There was no significant relationship among cell proliferation,apoptosis,clinical stages and pathological grades.High AI was always asso-ciated with a poor prognosis of the patients. Conclusion Cell proliferation and apoptosis allow to distinguish among normal epithelium,CIN and cervical carcinoma and are useful for the assessment of the malignant potential of tumor tissues.     

PARP inhibitor reduces proliferation and increases apoptosis in breast cancer cells

Objective： Apoptosis is a reliable marker of chemotherapeutic efficacy. Olaparib and paclitaxel inhibit proliferation and induce apoptosis in a variety of cancers. We investigated the effects of paclitaxel combined with olaparib on apoptosis in breast cancer Bcap37 cells. Methods： Proliferation and apoptosis were detected by MTT assay and PI staining. Degradation of procaspase-3 and poly（ADP-ribose） polymerase （PARP） was analyzed by Western blotting. Results： Compared with paclitaxel alone, paclitaxel combined with 100 mg olaparib significantly reduced survival in Bcap37 cells at all tested treatment durations （P〈0.05）; inhibition increased with increasing olaparib dose and treatment time （P〈0.01）. Combined treatment yielded significantly higher rates of apoptosis （P〈0.05）, which also increased with time （P〈0.01）. Fluorescence micrographs showed that early and late apoptotic cells increased with treatment time. Pro-caspase-3 and PARP degradation was induced by paclitaxel and enhanced by olaparib in a dose-dependent manner. Thus, combined treatment was substantially more effective than treatment with paclitaxel alone. Conclusions： Our findings suggest that paclitaxel and olaparib inhibit breast cancer Bcap37 cell proliferation and induce apoptosis. Combined treatment further reduced cell growth and enhanced apoptosis, suggesting that this combination therapy may be a promising treaunent for breast cancer.     

The effect and significance on Akt and PTEN expression in melanoma B16 cells with chamaejasme extract

Objective： The aim of this study was to investigate the effect on Akt and PTEN expression and the mechanism of proliferation and apoptosis of melanoma B16 cells treated with chamaejasme extract. Methods： The expressions of Akt and PTEN of B16 cells treated with different concentrations of chamaejasme extract were detected with immunohistochemical method, cell apoptosis index （AI） was calculated with in situ labeling method. Results： Akt expressions of B16 cells treated with different concentrations of chamaejasme extract were significantly lower than the control group, while the PTEN expres- sions significantly up-regulated, and the effects appeared to be dose-related （P 〈 0.05）. The Akt/AI of B16 cells treated with different concentrations of chamaejasme extract was significantly lower than the control group, all the parameters had ex- tremely difference （F = 24.58, P 〈 0.05）. Conclusion： Chamaejasme extract can inhibit proliferation and induce apoptosis of malignant melanoma B 16 cells by down-regulating the expression of Akt and up-regulating the expression of PTEN.     

Effects of Imbalance of Apoptosis and Proliferation on Large Bowel Carcinogenesis in Mice

OBJECTIVE To observe the pattern of changes in the proliferation and apoptosis at different stages of large bowel carcinoma in mice, and to explore the effects of the imbalance of apoptosis and proliferation at different stages of large-intestine carcinogenesis.METHODS An experimental animal model for large intestine carcinogenesis of KUNMING-strain mice was used. The carcinomas were induced by subcuteneous injection of dimethylhydrazine (DMH) and the distribution and density changes of proliferating and apoptotic cells observed through multistages toward cancer formation. The animals were killed in groups at the 12th, 18th, 24th,and 32nd weeks of carcinoma induction. The apoptotic and proliferating cells were labeled separately using TUNEL and PCNA immunohistochemical staining methodsRF, RESULTS In the normal mouse mucosa, all the apoptotic cells were situated in the superficial layers, however, the proliferating cells were situated in the basement layers, and the amount of both were small. In the early stage of carcinoma induction, the proliferation and the apoptotic cells slightly increased in amount, but there were no obvious changes in their ratio. In the medium stage, the densities of both distinctly increased, but there were no obvious changes in the ratio. In the late stage, the densities of the proliferating and the apoptotic cells in the non-carcinoma mucosa were higher than those at other stages. The proliferating cells in the dysplastic mucosa increased progressively with the increasing degree of the lesions. Although the apoptotic cells increased, their changes did not occur with the degree of the lesions. Their ratio showed a decreasing tendency with the degree of the lesions.CONCLUSIONS (①The presence of an imbalance between cell proliferation and apoptosis was confirmed in the course of large intestine carcinogenesis in a mouse model. ②In the early stage of carcinoma induction both proliferation and apoptosis were at a low level; in the medium stage, they were both at a high level; and in the late stage (that is in carcinoma), proliferation was at a very high level, while apoptosis was at a low level. ③The proliferating cells increased progressively with the degree of dysplasia. There were no obvious changes in the apoptotic cells and their ratio to the proliferating cell sshowed a progressively increasing tendency. ④In the stage of cancer formation, the most essential change was the excessive decrease in the ratio of apoptosis to proliferation. These results support the hypothesis of “Cell Selective Proliferation“, which was raised by authors previously in a study on human large bowel carcinoma.     

Effects of shRNA interfering hexokinaseII gene expression on the malignant biological behaviors of human lymphoma cells北大核心CSCD

Objective: To investigate the effects of hexokinaseII (HK II) gene-silencing by shRNA interference on cell metabolism, proliferation, apoptosis and drug-resistance of human lymphoma cell lines Raji and SU-DHL-4, therefore to evaluate the potential value of HK II gene-targeted therapy for the treatment of lymphoma. Methods: The lymphoma cell lines Raji and SU-DHL-4 transfected with lentiviral vectors carring shRNAs targeting HK II gene (HKII shRNA) were established, while the control vectors carring negative control shRNAs were used as the control group. The expression levels of HKII mRNA and protein were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. Then the cell viability was assessed using trypan blue staining, and the cell growth curve was drawn. The half maximal inhibitory concentration (IC50) of doxorubicin (DOX) for lymphoma cells was examined by CCK-8 method. The cell cycle distribution and apoptosis were analyzed by flow cytometry. The expression levels of apoptosis-related proteins caspase-3, cleaved caspase-3 (c-caspase-3), Bcl-2 and Bcl-6 were detected by Western blotting. The concentrations of lactic acid and glucose in the supernatant of cell culture were analyzed using Lactic Acid Detection Kit and Glucose (HK) Assay Kit, respectively. Results: Compared with the control group, the levels of HKII mRNA and protein were significantly decreased in HKII shRNA transfected Raji and SU-DHL-4 cells (all P < 0.05). After HK II gene-silencing by HKII shRNA interference, the proliferation of lymphoma Raji and SU-DHL-4 cells was significantly inhibited (both P < 0.01), the cell cycle was arrested in G0/G1 phase (both P < 0.05), and the apoptosis was induced (both P < 0.01). Compared with the control group, HK II gene-silencing reduced IC50 value of DOX in lymphoma Raji and SU-DHL-4 cells (both P < 0.05), inhibited glucose consumption (both P < 0.05), and decreased lactic acid generation (both P < 0.01). In HKII shRNA transfected Raji and SUDHL- 4 cells, the level of Bcl-2 protein was down-regulated (both P < 0.05), and the level of c-caspase-3 was up-regulated (both P < 0.05). When the lymphoma Raji and SU-DHL-4 cells were treated with DOX (0.1 μmol/L), the level of Bcl-2 in HKII shRNA transfection group was not significantly changed (both P > 0.05), whereas the level of caspase-3 was obviously decreased (both P < 0.01), and the level of c-caspase-3 was significantly increased (both P < 0.05) as compared with the control group. Conclusion: HK II gene-silencing by shRNA interference can suppress the proliferation, promote the apoptosis, correct the glycolytic metabolism phenotype, and enhance the sensitivity of lymphoma cells to chemotherapeutic drug DOX. It suggests that HK II gene may be a potential target for the treatment of relapsed aggressive lymphoma. Copyright © 2017 by TUMOR All rights reserved.     

Aberrant DNA methyltransferase 1 expression in clear cell renal cell carcinoma development and progression

Objective： To better understand the contribution of dysregulated DNA methyltransferase 1 （DNMT1） expression to the progression and biology of clear cell renal cell carcinoma （ccRCC）. Methods： We examined the differences in the expression of DNMT1 in 89 ecRCC and 22 normal tissue samples by immunohistochemistry. In addition, changes in cell viability, apoptosis, colony formation and invading ability of ccRCC cell lines （786-0 and Caki-1） were assessed after transfection with DNMT1 siRNA. Results： We found DNMT1 protein was significantly higher expressed in ccRCC than that of in no-tumor tissues （56.2% and 27.3%, respectively, P=0.018）. The expression of DNMT1 was strongly associated with ccRCC tumor size, tumor pathology stage, histological grading, lymph node metastasis, vascular invasion, recurrence and prognosis. Moreover, knockdown of DNMT1 expression significantly inhibited ccRCC cell viability, induced apoptosis, decreased colony formation and invading ability. Conclusions： Expression of DNMTI protein is increased in ccRCC tissues, and DNMT1 expression is associated with poor prognosis of patients. Experiments in vitro further showed DNMT1 played an essential role in proliferation and invasion of renal cancer cells. Moreover, targeting this enzyme could be a promising strategy for treating ccRCC, as evidenced by inhibited cell viability, increased apoptosis, decreased colony formation and invading ability.     

Study of Multi-directional Derivation of Cord Blood Mononuclear Cells and Observation of Killing Activity to MGC-803 Gastric Cancer Cell Strain In Vitro

Objective： To study multi-directional derivation of cord blood mononuclear cells to CD3AK, LAK and CIK cells as well as changes of killing activity to gastric cancer cell strain in vitro. Methods： CD3mAb and IL-2 were used to induce CD3AK cells, and IL-2 was used to induce LAK cells; IFN-γ was used in the beginning, then IL-1, CD3mAb and IL-2 were used to induce CIK cells after 24 h for observing amplification and analyzing their relationship. The phenotypes of the cultured CIK cells were analyzed by flow cytometry. Subsequently, by using MGC-803 gastric cancer cell strain as target cells, the killing activity of CD3AK, LAK and CIK cells was evaluated by using MTT method. Results： The amplification activity of CD3AK and CIK cells was all far higher than LAK cells （P〈0.05）. The amplification activity had no obvious difference between CIK cells and CD3AK cells at prophase, but that was far higher in CIK cells than CD3AK cells at about 20^th day （P〈0.05）. The flow cytometry revealed that the amount of CD3^＋ CD56^＋ cells, major effector cells after CIK cells being cultured was significantly increased （P〈0.05）, moreover, the amount of CD8^＋ cells was significantly increased as well （P〈0.05）. The killing activities of CD3AK and CIK cells to the MGC-803 gastric cancer cell strain were all significantly higher than LAK cells, while the killing activity of CIK cells was stronger than CD3AK cells （P〈0.05）. Conclusion： CIK cells have stronger amplification activity and killing activity, and can be taken as more effective killing cells applied to the tumor adoptive immunotherapy.     

肿瘤性贫血骨髓幼红细胞促红细胞生成素受体及fas抗原表达的临床研究

 目的　探讨骨髓幼红细胞上促红细胞生成素受体（EPOR）和 fas抗原的表达在肿瘤性贫血发病机制中的意义。方法　以转铁蛋白受体（CD71）和血型糖蛋白（GPA）作为红系标志,采用流式细胞术检测了骨髓红系细胞上EPOR 和fas抗原的表达。结果　与缺铁性贫血患者比较 ,CD71细胞和GPA细胞上EPOR的表达[（17.21±16.47）%、（20.17±13.44）%;（11.41±12.14）%、（14.49±9.77）%]差异无统计学意义（P＞0.05和P＞0.05）,EPOR的表达水平同HGB水平之间也无明显相关性（ r = 1.21,P＞0.05）。但fas抗原的表达[（21.21±8.14）%、（18.30±7.10）%]显著高于缺铁性贫血患者[（6.40±5.72）%、（8.71±6.55）%]（P＞0.05）,且fas抗原的表达与HGB的水平呈负相关（r = 0.71,P＜0.05）。结论　红系细胞凋亡率增高可能同肿瘤性贫血的发病机制有关, EPOR 表达水平降低可能不是肿瘤性贫血发病的主要机制。     

干细胞因子与促红细胞生成素联合应用对非霍奇金淋巴瘤贫血患者红系生成的作用

目的:探讨体外液体培养条件下干细胞因子(SCF)与促红细胞生成素(Epo)协同刺激对非霍奇金淋巴瘤(NHL)贫血患者骨髓红系造血的作用。方法:将NHL贫血患者骨髓单个核细胞(MNC)接种于液体培养基,在无细胞因子条件下和分别添加SCF、EPO及SCF EPO的刺激条件下培养7天,通过计算红系恢复性增生份数和流式细胞术评价红系的增生和分化。结果:SCF EPO刺激条件下的红系恢复性增生份数高于EPO单独刺激者,差异具有显著性意义(P<0.02)。Epo或SCF Epo刺激培养7天后的红系分化主要处于分化早期的原始红细胞阶段。流式细胞术显示红系分化以CD71阳性细胞为主。与单用Epo刺激相比,SCF Epo刺激培养后的CD71阳性红系细胞比率增高,差异具有显著性意义(P<0.01)。结论:在体外液体培养条件下,SCF可以增强Epo的促NHL贫血患者骨髓红系增生、分化作用,这为肿瘤性贫血的治疗提供了新的启示。     

Resveratrol induces human K562 cell apoptosis,erythroid differentiation,and autophagy

Resveratrol (Res) is a naturally occurring phytoalexin with apoptotic and inducing-glob effects in leukemic cells, but the potential induction of erythroid differentiation in cells is not fully understood. Here, we investigated the effects of Res on human erythro-megakaryoblastic leukemia cell line K562. Among the treated cells, proliferation was inhibited and the occurrence of cell apoptosis and cell death were detected. Erythroid differentiation assay was explored, and we found that Res could increase the expression of glycophorin A (GPA), HBA1, HBB, and γ-globin genes and enforced the expression of GPA, CD71, and Band3 proteins. Res also induced K562 cell autophagy when the concentration of Res was increased up to 50 or 100 μM. Our findings suggested that Res possesses the potency not only inducing apoptosis but also inducing erythroid differentiation and autophagy in K562 cells. These results provide that Res may be a therapeutic candidate for chronic myelogenous leukemia treatment.     

CGI-100 specific shRNA inhibits proliferation and induces differentiation in leukemia K562 cells

Objective: To investigate the characteristics of CGI-100- knockdown K562 cells and the effect of CGI-100 RNA interference (RNAi) on matrine-treated K562 cells.Methods: Three oligonucleotides targeting CGI-100 gene and a pair of negative control containing the same nucleotide composition with a different sequence were devised and chemically synthesized. The inhibition efficiency of CGI-100 expression by shRNA-CGI-100 in K562 cells was determined using semiquantitative RT-PCR and dot blot hybridization. The effect of CGI-100 RNAi on the growth of K562 cells was examined using MTT assay and cell differentiation was measured by distinct approaches including flow cytometry, benzidine staining and electron microscope. After CGI-100-konckdown K562 cells were incubated with 0.2 mg/ml of matrine or 30 μmol/L of hemin for 48 h, the expression levels of Glycophorin A(GPA)(CD235a) and Growth factor independence-1B mRNA(Gfi-1B mRNA) were measured by RT-PCR and the protein levels of GPA, CD14 and CD15 were detected by flow cytometry.Results: The eukaryotic expression vectors of CGI-100 RNAi were successfully constructed. The K562/shRNA-CGI-100 cell line was established in which the inhibition efficiency of CGI-100 gene expression by shRNA-CGI-100 was 54%. CGI-100-knockdown inhibited the proliferation and induced erythroid differentiation in K562 cells. Compared with the control K562 ceils, the K562/shRNA-CGI-100 cells showed decreased absorbance value detected by MTT assay, decreased enchromation, increased heterochromation, increased percentage of G0/G>1 phase cells, decreased population of S phase cells, decreased PI (proliferation index of cells), and elevated percentage of benzidine-positive cells. Moreover, the sensitivity of K562/shRNA-CGI-100 cells to either matrine or hemin was enhanced and the sensitivity of these cells to matrine was higher than that to hemin. Compared with the control K562 cells, matrine treatment in K562/shRNA-CGI-100 cells resulted in increased inhibitory rate of proliferation, elevated percentage of benzidine-positive cells, obviously up-regulated mRNA expressions of GPA and Gfi-1B, and increased mean fluorescence intensity (MFI) of GPA. No CD14 expression was detected and no statistical significance was found for the detected CD15. Finally, the MFI of GPA increased in K562/shRNA-CGI-100 cells treated with hemin and was 1.7 times less than that in cells exposed to matrine.Conclusion: These results suggest that the function of CGI-100 gene is correlated with the deregulated proliferation and the block of erythroid differentiation in K562 cells and may also be involved in matrine-induced erythroid differentiation in K562 cells.     

Homing-associated cell adhesion molecule (H-CAM/CD44) on human CD34+ hematopoietic progenitor cells.

Human CD34+ hematopoietic progenitor cells (HPCs) express CD44 and can directly adhere to hyaluronate (HA) via CD44. Furthermore, CD44 may also be involved in the regulation of CD34+ HPC proliferation and development. The expression of CD44 molecules on CD34+ hematopoietic progenitor cells is significantly lower on bone marrow (BM) CD34+ cells compared with circulating CD34+ cells in cord blood and peripheral blood. Myeloid and erythroid progenitor cells are found predominantly in CD34+ CD44+ cell fractions. More interestingly, CD34+ CD44- cells expressing B-lymphocyte-associated CD10 and CD19 would represent unique B-lymphocyte committed precursors in the BM, which might undergo apoptotic cell death in the early steps of B-cell differentiation.     

Comparative analysis of the in vitro proliferation and expansion of hematopoietic progenitors from patients with aplastic anemia and myelodysplasia

Aplastic anemia (AA) and myelodysplasia (MDS) show great similarities in their biology. To date, however, it is still unclear to what extent hematopoietic progenitor cells (HPCs) from AA and MDS share biological properties and what the functional differences are between them. In trying to address this issue, in the present study we have analyzed, in a comparative manner, the proliferation and expansion capacities of bone marrow (BM) progenitor cells from AA and MDS in response to recombinant cytokines. BM samples from normal subjects (NBM) and patients with AA and MDS were enriched for HPC by immunomagnetic-based negative selection. Selected cells were cultured in the absence (control) or in the presence of early-acting cytokines (Mix I), or early-, intermediate- and late-acting cytokines (Mix II). Proliferation and expansion were assessed periodically. In NBM and MDS cultures apoptosis was also determined. In NBM cultures, Mix I induced a nine-fold increase in total cell numbers and a 3.6-fold increase in colony-forming cell (CFC) numbers. In Mix II-supplemented cultures, total cells were increased 643-fold, and CFC 12.4-fold. In AA cultures, no proliferation or expansion were observed in Mix I-supplemented cultures, whereas only a four-fold increase in total cell numbers was observed in the presence of Mix II. In MDS cultures, a 12-fold increase in total cells and a 2.9-fold increase in CFC were observed in the presence of Mix I; on the other hand, Mix II induced a 224-fold increase in total cells and a 5.9-fold increase in CFC. Apoptosis was reduced in cytokine-supplemented cultures from NBM. In contrast, Mix II induced a significant increase in the rate of apoptosis in MDS cultures. Our results demonstrate that, as compared to their normal counterparts, AA and MDS progenitors are deficient in their proliferation and expansion potentials. Such a deficiency is clearly more pronounced in AA cells, which seem to be unable to respond to several cytokines. MDS progenitors, on the other hand, are capable to proliferate and expand in response to cytokines; however, their rate of apoptosis is increased by intermediate- and late-acting cytokines, so that the overall proliferation and expansion are significantly lower than those of normal progenitor cells.     

Laptm53′UTR对小鼠B细胞淋巴瘤38B9细胞增殖与凋亡的影响

目的:通过建立稳定过表达溶酶体相关穿膜蛋白5(lysosomal associated protein transmembrane 5,Laptm5)3'不翻译区(3'UTR)的B细胞淋巴瘤38B9细胞株,探讨Laptm5 3'UTR对小鼠B细胞淋巴瘤38B9细胞株增殖、凋亡的影响.方法:荧光定量PCR及Western blotting检测小鼠B细胞及38B9细胞中Laptm5 miRNA及其蛋白质的表达水平;将小鼠Laptm5 3'UTR及其含有的microRNA结合位点突变的突变型基因片段构建入逆转录病毒表达载体pMSCV-PIG,包装成逆转录病毒,感染小鼠B细胞淋巴瘤细胞38B9,流式细胞术检测逆转录病毒感染率,细胞计数法及流式细胞术观察Laptm53'UTR或其突变型过表达后389B9细胞的增殖、凋亡情况.结果:相比小鼠B细胞,B细胞淋巴瘤细胞中Laptm5的miRNA及蛋白均显著降低(P＜0.01).成功获得稳定过表达Laptm5 3'UTR(突变型)的38B9细胞株.Laptm5 3'UTR细胞株比Laptm5 3'UTR突变型过表达细胞株的增殖能力显著减慢,凋亡率显著增加[(7.87±1.08)％vs(0.45±0.07)％,P＜0.01].结论:小鼠Laptm5 3'UTR具有抑制B细胞淋巴瘤增殖、促进其凋亡的作用,该作用可能与其影响相关microRNA的调控有关.     

Homing-Associated Cell Adhesion Molecule (H-CAM/CD44) on Human CD34+ Hematopoietic Progenitor Cells

Human CD34⁺ hematopoietic progenitor cells (HPCs) express CD44 and can directly adhere to hyaluronate (HA) via CD44. Furthermore, CD44 may also be involved in the regulation of CD34⁺ HPC proliferation and development. The expression of CD44 molecules on CD34⁺ hematopoietic progenitor cells is significantly lower on bone marrow (BM) CD34⁺ cells compared with circulating CD34⁺ cells in cord blood and peripheral blood. Myeloid and erythroid progenitor cells are found predominantly in CD34⁺CD44^- cell fractions. More interestingly, CD34⁺CD44⁺ cells expressing B-lymphocyte-associated CD10 and CD19 would represent unique B-lymphocyte committed precursors in the BM, which might undergo apoptotic cell death in the early steps of B-cell differentiation.     

Antiproliferative and apoptotic effects of mycophenolic acid in human B-cell non-Hodgkin lymphomas

Mycophenolic acid (MPA)/mycophenolate mofetil (MMF), a powerful immunosuppressive agent was tested on human B-lymphoma cells (Epstein-Barr virus +/-) in vitro and in SCID mouse xenograft model. Proliferation, apoptotic activity and tumor volume were evaluated. MPA inhibited lymphoma cell proliferation and induced apoptosis (50-60% at 72 h). In vivo, oral administration significantly inhibited subcutaneous tumor growth. Immunohistochemistry showed significantly decreased proliferation rate and higher apoptotic activity in tumors treated with MMF. Xenografted lymphoma cells remained sensitive to MPA. Our results suggest that MPA may be recommended as an additional component of lymphoma chemotherapeutical regimens, with special considerations to post-transplant lymphomas.     

Granulocyte colony-stimulating factor inhibits Fas-triggered apoptosis in bone marrow cells isolated from patients with refractory anemia with ringed sideroblasts.

Treatment with granulocyte colony-stimulating factor (G-CSF) plus erythropoietin may synergistically improve hemoglobin levels and reduce bone marrow apoptosis in patients with refractory anemia with ringed sideroblasts (RARS). Fas-induced caspase activity is increased in RARS bone marrow cells. We showed that G-CSF significantly reduced Fas-mediated caspase-8 and caspase-3-like activity and the degree of nuclear apoptotic changes in bone marrow from nine RARS patients. A decrease in mitochondrial membrane potential and an increase in intracellular reactive oxygen species occurred in Fas-treated cells, but became significant only 24 h after changes in caspase activity and decrease in proliferation. G-CSF also reduced the magnitude of these late apoptotic changes. In CD34-selected normal cells, G-CSF induced myeloid colony growth, and an overall small decrease in the number of erythroid colonies. By contrast, G-CSF induced a 33-263% increase of erythroid colony formation in CD34+ cells from four of five RARS patients with severely reduced erythroid growth, while the normal or slightly reduced erythroid growth of three other patients was not influenced by G-CSF. This study suggests that G-CSF may reduce the pathologically increased caspase activity and concomitant apoptotic changes, and promote erythroid growth and differentiation of stem cells from RARS patients. Our data support the clinical benefit of G-CSF in this subgroup of myelodysplastic syndromes.