Studies on the Antimitotic Activity of Leurocristine (Vincristine)

Abstract The effect of leurocristine (vincristine) on cell division was studied in normal bone marrow and leukemic cells (L1210 leukemia) of DBA/2 mice. It was found that vincristine possesses the ability to arrest mitosis at the metaphase stage. The accumulation of arrested metaphases in the normal bone marrow and in leukemic cells of the animals treated with vincristine followed a pattern similar to the one observed after treatment with colchicine or vincaleukoblastine (vinblastine).

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The Catharanthus Roseus (Vinca Rosea) Alkaloids: A New Glass of Stathmokinetic Agents

Acta geneticae medicae et gemellologiae ◽

10.1017/s1120962300026445 ◽

1968 ◽

Vol 17 (1) ◽

pp. 197-208 ◽

Cited By ~ 1

Author(s):

G. Cardinali ◽

G. Centurelli

Keyword(s):

Bone Marrow ◽

Antitumor Activity ◽

Catharanthus Roseus ◽

Biological Activities ◽

Leukemic Cells ◽

Normal Bone Marrow ◽

Normal Bone ◽

Vinca Rosea ◽

Antimitotic Activity ◽

The Relationship

SummaryVinblastine (VLB) and vincristine (VCR) are two alkaloids (extracted from the plant Catahranthus roseus) which possess a definite antitumor activity. The antitumor activity. The antimitotic action of these two alkaloids was studied both on normal bone marrow and leukemic cells. Comparative studies showed that they are, like colchicine, spindle poisons. They, however, differ from colchicine and its derivatives in some of their biological activities.The problem of the relationship between antitumor and antimitotic activity of VLB and VCR is discussed.

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Expression of the three myeloid cell-associated immunoglobulin G Fc receptors defined by murine monoclonal antibodies on normal bone marrow and acute leukemia cells

Blood ◽

10.1182/blood.v73.7.1951.bloodjournal7371951 ◽

1989 ◽

Vol 73 (7) ◽

pp. 1951-1956

Author(s):

ED Ball ◽

J McDermott ◽

JD Griffin ◽

FR Davey ◽

R Davis ◽

...

Keyword(s):

Bone Marrow ◽

Monoclonal Antibodies ◽

Acute Leukemia ◽

Cell Sorting ◽

Lymphoblastic Leukemia ◽

Mononuclear Phagocytes ◽

Leukemic Cells ◽

Leukemia Cells ◽

Normal Bone Marrow ◽

Normal Bone

Monoclonal antibodies (MoAbs) have been prepared recently that recognize the three cell-surface receptors for the Fc portion of immunoglobulin (Ig), termed Fc gamma RI (MoAb 32.2), Fc gamma R II (MoAb IV-3), and Fc gamma R III (MoAb 3G8) that are expressed on selected subsets of non-T lymphocyte peripheral blood leukocytes. In the blood, Fc gamma R I is expressed exclusively on monocytes and macrophages, Fc gamma R II on granulocytes, mononuclear phagocytes, platelets, and B cells, and Fc gamma R III on granulocytes and natural killer (NK) cells. We have examined the expression of these molecules on normal bone marrow (BM) cells and on leukemia cells from the blood and/or BM in order to determine their normal ontogeny as well as their distribution on leukemic cells. BM was obtained from six normal volunteers and from 170 patients with newly diagnosed acute leukemia. Normal BM cells were found to express Fc gamma R I, II, and III with the following percentages: 40%, 58%, and 56%, respectively. Cell sorting revealed that both Fc gamma R I and Fc gamma R II were detectable on all subclasses of myeloid precursors as early as myeloblasts. Cell sorting experiments revealed that 66% of the granulocyte-monocyte colony-forming cells (CFU-GM) and 50% of erythroid burst-forming units (BFU-E) were Fc gamma R II positive with only 20% and 28%, respectively, of CFU-GM and BFU-E were Fc gamma R I positive. Acute myeloid leukemia (AML) cells expressed the three receptors with the following frequency (n = 146): Fc gamma R I, 58%; Fc gamma R II, 67%; and Fc gamma R III, 26% of patients. Despite the fact that Fc gamma R I is only expressed on monocytes among blood cells, AML cells without monocytoid differentiation (French-American-British [FAB]M1, M2, M3, M6) were sometimes positive for this receptor. However, Fc gamma R I was highly correlated with FAB M4 and M5 morphology (P less than .001). Fc gamma R II was also correlated with FAB M4 and M5 morphology (P = .003). Cells from 11 patients with acute lymphoblastic leukemia were negative for Fc gamma R I, but six cases were positive for Fc gamma R II and III (not the same patients). These studies demonstrate that Ig Fc gamma R are acquired during normal differentiation in the BM at or before the level of colony-forming units. In addition, we show that acute leukemia cells commonly express Fc gamma R.

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Expression of the three myeloid cell-associated immunoglobulin G Fc receptors defined by murine monoclonal antibodies on normal bone marrow and acute leukemia cells

Blood ◽

10.1182/blood.v73.7.1951.1951 ◽

1989 ◽

Vol 73 (7) ◽

pp. 1951-1956 ◽

Cited By ~ 27

Author(s):

ED Ball ◽

J McDermott ◽

JD Griffin ◽

FR Davey ◽

R Davis ◽

...

Keyword(s):

Bone Marrow ◽

Monoclonal Antibodies ◽

Acute Leukemia ◽

Cell Sorting ◽

Lymphoblastic Leukemia ◽

Mononuclear Phagocytes ◽

Leukemic Cells ◽

Leukemia Cells ◽

Normal Bone Marrow ◽

Normal Bone

Abstract Monoclonal antibodies (MoAbs) have been prepared recently that recognize the three cell-surface receptors for the Fc portion of immunoglobulin (Ig), termed Fc gamma RI (MoAb 32.2), Fc gamma R II (MoAb IV-3), and Fc gamma R III (MoAb 3G8) that are expressed on selected subsets of non-T lymphocyte peripheral blood leukocytes. In the blood, Fc gamma R I is expressed exclusively on monocytes and macrophages, Fc gamma R II on granulocytes, mononuclear phagocytes, platelets, and B cells, and Fc gamma R III on granulocytes and natural killer (NK) cells. We have examined the expression of these molecules on normal bone marrow (BM) cells and on leukemia cells from the blood and/or BM in order to determine their normal ontogeny as well as their distribution on leukemic cells. BM was obtained from six normal volunteers and from 170 patients with newly diagnosed acute leukemia. Normal BM cells were found to express Fc gamma R I, II, and III with the following percentages: 40%, 58%, and 56%, respectively. Cell sorting revealed that both Fc gamma R I and Fc gamma R II were detectable on all subclasses of myeloid precursors as early as myeloblasts. Cell sorting experiments revealed that 66% of the granulocyte-monocyte colony-forming cells (CFU-GM) and 50% of erythroid burst-forming units (BFU-E) were Fc gamma R II positive with only 20% and 28%, respectively, of CFU-GM and BFU-E were Fc gamma R I positive. Acute myeloid leukemia (AML) cells expressed the three receptors with the following frequency (n = 146): Fc gamma R I, 58%; Fc gamma R II, 67%; and Fc gamma R III, 26% of patients. Despite the fact that Fc gamma R I is only expressed on monocytes among blood cells, AML cells without monocytoid differentiation (French-American-British [FAB]M1, M2, M3, M6) were sometimes positive for this receptor. However, Fc gamma R I was highly correlated with FAB M4 and M5 morphology (P less than .001). Fc gamma R II was also correlated with FAB M4 and M5 morphology (P = .003). Cells from 11 patients with acute lymphoblastic leukemia were negative for Fc gamma R I, but six cases were positive for Fc gamma R II and III (not the same patients). These studies demonstrate that Ig Fc gamma R are acquired during normal differentiation in the BM at or before the level of colony-forming units. In addition, we show that acute leukemia cells commonly express Fc gamma R.

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Loss of suppression of normal bone marrow colony formation by leukemic cell lines after differentiation is induced by chemical agents

Blood ◽

10.1182/blood.v65.1.100.100 ◽

1985 ◽

Vol 65 (1) ◽

pp. 100-106 ◽

Cited By ~ 9

Author(s):

HN Steinberg ◽

AS Tsiftsoglou ◽

SH Robinson

Keyword(s):

Bone Marrow ◽

Cell Lines ◽

Colony Formation ◽

Leukemic Cell ◽

K562 Cells ◽

Leukemic Cells ◽

Cell Phenotype ◽

Normal Bone Marrow ◽

Normal Bone ◽

Leukemic Cell Lines

Abstract The human leukemic cell lines K562 and HL-60 were cocultured with normal bone marrow (BM) cells. Coculture with 10(4) K562 or HL-60 cells results in 50% inhibition of normal CFU-E and BFU-E colony formation. However, when the same number of K562 and HL-60 cells is first treated for two to five days with agents that induce their differentiation, a gradual loss in their capacity to inhibit CFU-E and BFU-E colony formation is observed. The inhibitory material in K562 cells is soluble and present in conditioned medium from cultures of these cells. The degree to which leukemic cell suppression of CFU-E and BFU-E growth is reversed is correlated with the time of exposure to the inducing agent. Suppression is no longer evident after five days of prior treatment with inducers. In fact, up to a 90% stimulation of CFU-E growth is observed in cocultures with K562 cells that have been pretreated with 30 to 70 mumol/L hemin for five days. K562 cells treated with concentrations of hemin as low as 30 mumol/L demonstrate increased hemoglobin synthesis and grow normally, but no longer have an inhibitory effect on CFU-E growth. Hence, reversal of normal BM growth inhibition must be caused by the more differentiated state of the K562 cells and not by a decrease in the number of these cells with treatment. Thus, induction of differentiation in cultured leukemic cells not only alters the malignant cell phenotype but also permits improved growth of accompanying normal marrow progenitor cells. Both are desired effects of chemotherapy.

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Novel Multiparametric Quantitative Immunocytomorphological Characterisation of Human Bone Marrow Precursor Differentiation and of Haematopoietic Neoplasms Using ImageStream® Cytometry

Blood ◽

10.1182/blood.v112.11.4871.4871 ◽

2008 ◽

Vol 112 (11) ◽

pp. 4871-4871

Author(s):

Catherine Claude Martin ◽

Chantal Jayat-Vignoles ◽

Jean-Luc Faucher ◽

Thaddeus George ◽

Vidya Venkatachalam ◽

...

Keyword(s):

Bone Marrow ◽

Flow Cytometry ◽

Blood Cells ◽

High Speed ◽

Tumour Cells ◽

Leukemic Cells ◽

Normal Bone Marrow ◽

Normal Cells ◽

Normal Bone ◽

Abnormal Cells

Abstract The ImageStream technology performs high speed acquisition of brightfield, laser scatter and up to four fluorescent images per cell for several thousands of cells in suspension, thereby enabling simultaneous immunophenotyping and morphology-based measurements. This is the only technology combining cytology and flow cytometry in one single platform. Our aim was to study normal and tumour cells of the haematopoietic lineage with this new technology in order to improve diagnosis of haematological disorders. We have defined cytomorphological criteria of normal bone marrow (n=4) and circulating blood cells (n=40). Cells were multi-colour labelled with both DRAQ5 nuclear stain and CD45 ECD-mAb, and additionally labeled with a combination of mAbs against either CD3/CD19, CD11b/CD16, CD14/CRTH2, or CD71/CD235. Results for normal cells were compared to those obtained by classical cytometry and cytology. We then applied these criteria to samples with patients with circulating leukemic cells, including 1 myelodysplatic syndrome (MDS), 1 myeloproliferative syndrome (MPS), 3 acute lymphoblastic leukaemia (ALL), 2 follicular lymphomas (FL) and 20 chronic lymphocytic lymphomas (CLL). We have created completely new quantitative cytomorphological criteria for classifying blood cells using parameters that measure cellular size and shape, nuclear to cytoplasmic area ratio, nuclear lobe count, SSC texture, the ratio between the size and the major axis of CD45, the ratio between the intensity and the compactness of SSC signal, and the intensity of DRAQ5 labelling, to name a few. Using these criteria, we have characterised normal bone marrow differentiation and normal circulating blood cells. We have obtained a perfect correlation with classical cytology and flow cytometry. Analysis of pathological samples showed that abnormal cells were recognized in all cases. We found an abnormal blast cell compartment and an abnormal monocytic differentiation branch in the case of MDS. We have also defined specific cytomorphological properties that distinguish ALL, FL and CLL tumour cells from normal cells. We also provide data that enumerates the proportion of large cells, of atypical CLL cells and of cells in the G2/M phase. Altogether, these results show that a technology combining cytology and flow cytometry in a single platform leads to the discovery of completely new and quantitative cytomorphological parameters defining each stage of normal cell and each category of abnormal cells of the haematopoietic lineage, opening completely new perspectives for the diagnosis of haematopoietic neoplasms.

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Loss of suppression of normal bone marrow colony formation by leukemic cell lines after differentiation is induced by chemical agents

Blood ◽

10.1182/blood.v65.1.100.bloodjournal651100 ◽

1985 ◽

Vol 65 (1) ◽

pp. 100-106

Author(s):

HN Steinberg ◽

AS Tsiftsoglou ◽

SH Robinson

Keyword(s):

Bone Marrow ◽

Cell Lines ◽

Colony Formation ◽

Leukemic Cell ◽

K562 Cells ◽

Leukemic Cells ◽

Cell Phenotype ◽

Normal Bone Marrow ◽

Normal Bone ◽

Leukemic Cell Lines

The human leukemic cell lines K562 and HL-60 were cocultured with normal bone marrow (BM) cells. Coculture with 10(4) K562 or HL-60 cells results in 50% inhibition of normal CFU-E and BFU-E colony formation. However, when the same number of K562 and HL-60 cells is first treated for two to five days with agents that induce their differentiation, a gradual loss in their capacity to inhibit CFU-E and BFU-E colony formation is observed. The inhibitory material in K562 cells is soluble and present in conditioned medium from cultures of these cells. The degree to which leukemic cell suppression of CFU-E and BFU-E growth is reversed is correlated with the time of exposure to the inducing agent. Suppression is no longer evident after five days of prior treatment with inducers. In fact, up to a 90% stimulation of CFU-E growth is observed in cocultures with K562 cells that have been pretreated with 30 to 70 mumol/L hemin for five days. K562 cells treated with concentrations of hemin as low as 30 mumol/L demonstrate increased hemoglobin synthesis and grow normally, but no longer have an inhibitory effect on CFU-E growth. Hence, reversal of normal BM growth inhibition must be caused by the more differentiated state of the K562 cells and not by a decrease in the number of these cells with treatment. Thus, induction of differentiation in cultured leukemic cells not only alters the malignant cell phenotype but also permits improved growth of accompanying normal marrow progenitor cells. Both are desired effects of chemotherapy.

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MYBL2 Is a Candidate Tumor Suppressor Gene In MDS

Blood ◽

10.1182/blood.v116.21.1865.1865 ◽

2010 ◽

Vol 116 (21) ◽

pp. 1865-1865

Author(s):

Stefan Heinrichs ◽

Lily Conover ◽

Carlos E. Bueso-Ramos ◽

Outi Kilpivaara ◽

Ross Levine ◽

...

Keyword(s):

Bone Marrow ◽

Cell Division ◽

Tumor Suppressor ◽

Bone Marrow Cells ◽

Normal Karyotype ◽

Differentially Expressed ◽

Normal Bone Marrow ◽

Normal Bone ◽

Cd34 Cells ◽

Marrow Cells

Abstract Abstract 1865 Cytogenetic changes, mainly deletions, can be found in about 30–50% of patients with Myelodysplastic Syndromes (MDS). To identify a tumor suppressor candidate within a commonly deleted region on chromosome 20q, we performed gene expression analysis on CD34+ bone marrow cells obtained from 8 patients with a 20q aberration and 18 with a normal karyotype. However, we were unable to identify genes that were significantly differentially expressed in aberrant 20q karyotype as compared to normal karyotype MDS patients. In contrast, a comparison of CD34+ cells from all MDS cases analyzed (n=26) with CD34+ cells obtained from normal bone marrow (n=4) revealed 108 genes that were differentially expressed. Interestingly, one of the top-scoring genes was MYBL2, which is located on chromosome 20q. MYBL2 levels were downregulated more than two-fold in 18 out of 26 cases. RNAi-mediated knockdown of MYBL2 in CD34+ normal bone marrow cells revealed a signature of genes functionally associated with the G2/M cell cycle phase confirming the well-documented role of MYBL2 as key transcription factor governing the onset of cell division. We hypothesize that in a subset of MDS cases the control of cell division may be impaired by low levels of MYBL2 such that altered cell fates established during cell division in early hematopoietic stem and progenitor cells will lead to clonal expansion with imbalanced or impaired differentiation. Indeed, gene set enrichment analysis revealed a strong enrichment of MYBL2 signature genes in MDS CD34+ cells. In support of a potential role as tumor suppressor, resequencing of MYBL2 (144 patients) identified 2 somatic mutations, pinpointing an additional mechanism to reduce expression of normal levels of wild-type MYBL2. Disclosures: No relevant conflicts of interest to declare.

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Selective regulation of the activity of different hematopoietic regulatory proteins by transforming growth factor beta 1 in normal and leukemic myeloid cells

Blood ◽

10.1182/blood.v76.7.1315.bloodjournal7671315 ◽

1990 ◽

Vol 76 (7) ◽

pp. 1315-1322 ◽

Cited By ~ 1

Author(s):

J Lotem ◽

L Sachs

Keyword(s):

Bone Marrow ◽

Transforming Growth Factor Beta ◽

Transforming Growth Factor ◽

Leukemic Cells ◽

Normal Bone Marrow ◽

Tgf Beta ◽

Gm Csf ◽

Normal Bone ◽

Growth Factor Beta ◽

Cells Cultured

The viability of normal bone marrow myeloid precursor cells induced by interleukin-6 (IL-6) or IL-1 alpha and the ability of IL-6 and IL-1 alpha to induce the formation of colonies of granulocytes, macrophages, or megakaryocytes in densely seeded bone marrow cultures was suppressed by transforming growth factor-beta 1 (TGF-beta 1). Induction of normal bone marrow colony formation by IL-3 was much less sensitive to TGF- beta 1, and there was little or no effect of TGF-beta 1 on colony formation induced by macrophage colony-stimulating factor (M-CSF) or granulocyte-macrophage CSF (GM-CSF). In different clones of myeloid leukemic cells, TGF-beta 1 suppressed differentiation induced with IL- 6, IL-1 alpha, or lipopolysaccharide (LPS), but did not suppress differentiation induced with IL-3 or GM-CSF. The effect of TGF-beta 1 on differentiation of the leukemic cells can be dissociated from its effect on cell growth. TGF-beta 1 suppressed the production of IL-6 in normal bone marrow cells cultured with IL-1 alpha and the production of IL-6 and GM-CSF in leukemic cells cultured with IL-1 alpha or LPS. The suppression of IL-6 production can explain the suppression by TGF-beta 1 of the effects of IL-1 alpha and LPS that are mediated by IL-6. TGF- beta 1 also suppressed differentiation in clones of myeloid leukemic cells induced with differentiation factor/leukemia inhibitory factor and tumor necrosis factor. In different leukemic clones TGF-beta 1 suppressed or enhanced induction of differentiation with dexamethasone. The results show that TGF-beta 1 can selectively control the activity of different molecular regulators of normal and leukemic hematopoiesis.

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Inhibition of human bone marrow fibroblast colony formation by leukemic cells

Blood ◽

10.1182/blood.v62.6.1261.bloodjournal6261261 ◽

1983 ◽

Vol 62 (6) ◽

pp. 1261-1265 ◽

Cited By ~ 1

Author(s):

T Nagao ◽

K Yamauchi ◽

M Komatsuda ◽

K Noguchi ◽

M Shimizu ◽

...

Keyword(s):

Bone Marrow ◽

Colony Formation ◽

Bone Marrow Cells ◽

Culture Media ◽

Suppressive Effect ◽

Leukemic Cells ◽

Normal Bone Marrow ◽

Humoral Factors ◽

Normal Bone ◽

Marrow Cells

The number of bone marrow fibroblast colony formations decreases in most cases of acute leukemia before the initiation of chemotherapy. This study was undertaken to clarify the mechanism of suppression of fibroblast colony formation in leukemic patients. Titration of the number of bone marrow cells did not indicate a linear relationship between the number of bone marrow cells cultured and the number of fibroblast colony formations. The number of fibroblast colony formations recovered by removal of nonadherent leukemic cells following one day of incubation increased. The cloning efficiency of patient bone marrow still showed increases in colony formation at higher plating concentrations after the nonadherent cells were removed. When leukemic and normal bone marrow cells were cocultured, the suppressive effect of leukemic cells on normal marrow fibroblast colony formation was clearly observed. The suppressive effect disappeared at complete remission, and then reappeared at relapse. Heat-inactivated serum and bone marrow culture media from leukemic patients whose fibroblast colony formations were small in number suppressed fibroblast colony formation from normal bone marrow. From these results, it was concluded that the suppression of fibroblast colony formation in leukemic patients was through humoral factors produced by leukemic cells.

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Improved Survival of Leukemic Mice Treated with Sodium Caseinate in Combination with Daunorubicin without Toxicity

Journal of Oncology ◽

10.1155/2021/6635650 ◽

2021 ◽

Vol 2021 ◽

pp. 1-8

Author(s):

Itzen Aguiñiga-Sánchez ◽

Frida Montserrat Meléndez-Ibarra ◽

Edgar Ledesma-Martínez ◽

Benny Weiss-Steider ◽

Guadalupe Rosario Fajardo-Orduña ◽

...

Keyword(s):

Bone Marrow ◽

Survival Rate ◽

Mononuclear Cells ◽

Bone Marrow Cells ◽

Leukemia Cell ◽

Sodium Caseinate ◽

Leukemic Cells ◽

Normal Bone Marrow ◽

Normal Bone ◽

Marrow Cells

In recent years, low doses of chemotherapy have been resumed and explored for the treatment of acute myeloid leukemia. Thus, CPX-351, a dual-drug liposomal encapsulation of cytarabine and daunorubicin, was approved by the US Food and Drug Administration, to deliver a synergistic 5 : 1 molar drug ratio into leukemia cells to a greater extent than normal bone marrow cells and significantly enhance survival compared with conventional treatment in older and newly diagnosed AML patients, but overall survival rate remains low; therefore, the need for new therapeutic options continues. Sodium caseinate (SC), a salt of casein, the main milk protein, has cytotoxic effect in leukemia cell lines, but promotes proliferation of hematopoietic normal cells, while its administration in leukemic mice promotes survival for more than 40 days, but bone marrow surviving mice still harbour leukemic cells, but it is not known whether the combination with cytarabine or daunorubicin can improve survival without damaging normal hematopoietic cells. Here, it is shown that, in vitro, the combination of the IC25 of SC-cytarabine or SC-daunorubicin synergizes in the elimination of leukemic cells, with evident induction of apoptosis, while the proliferation of mononuclear cells of bone marrow is not affected. In leukemic mice, the combined administration of SC-daunorubicin or SC-cytarabine promotes the highest survival rate at 40 days; in addition, no autoproliferating cells were detected in the bone marrow of survivors of more than 60 days, evidence of eradication of leukemic cells, but only the bone marrow of mice treated with the SC-daunorubicin combination proliferated in the presence of interleukin-3, which shows that this combination is not toxic to normal bone marrow cells, thus emerging as a possible antileukemic agent.

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