CML患者のbcr/abl(+)-CD34+細胞の増殖と分化に及ぼすG-CSFの効果

[Effect of G-CSF on the proliferation and differentiation of bcr/abl(+)-CD34+ cells from CML patients].

• Cheng-Yu Li
• Fan-Yi Meng
• Qi-Xin Sun
• Yun-Bi Fu
• Qian-Li Jiang
• Zheng-Shan Yi
• Lan-Lin Song

抄録

目的:

顆粒球コロニー刺激因子（G-CSF）がbcr/abl(+)-CD34+細胞の増殖と分化に及ぼす影響を調べる。

OBJECTIVE: To study the effect of granulocyte colony-stimulating factor (G-CSF) on the proliferation and differentiation of bcr/abl(+)-CD34+ cells.

方法:

慢性骨髄性白血病（CML）患者の骨髄からbcr/abl(+)-CD34+細胞を分離し、0, 10, 100, 1000 ng/mlのG-CSFを48, 96, 144時間かけて投与した。正常骨髄からのCD34細胞をコントロールとして使用した。細胞増殖はトリパンブルー色素排除法で決定し、細胞周期と抗原分化はフローサイトメトリーで決定し、細胞形態は光顕微鏡下で観察した。

METHODS: bcr/abl(+)-CD34+ cells were isolated from bone marrow of chronic myelocytic leukemia (CML) patients and were treated with 0, 10, 100, 1000 ng/ml of G-CSF for 48, 96, 144 hs. CD34 cells from normal bone marrow were used as controls. Cell proliferation was determined by trypan blue dye exclusion, cell-cycle and antigen differentiation were determined by flow cytometry and cell morphology was observed under light microscope.

結果:

bcr/abl(+)-CD34+細胞数はすべての群で明らかに増加した。48時間および96時間培養後、G-CSF 10 ng/ml群のbcr/abl(+)-CD34+細胞数は、G-CSF 0 ng/ml群に比べて有意に増加した（P < 0.05）が、正常なCD34細胞数はG-CSFの存在下でのみ増加した。48、96および144時間培養後、G-CSF 100 ng/ml群の細胞数は、G-CSF 0 ng/ml群に比べて有意に増加した（それぞれP < 0.05、P < 0.01、P < 0.01、P < 0.01）。144時間培養後、G-CSF 10, 100, 1000 ng/ml群のbcr/abl(+)-CD34+細胞のG0/G1期における細胞割合はG-CSF 0 ng/ml群に比べて有意に少なく（P < 0.05）、G-CSF 10, 100, 1000 ng/ml群の正常CD34細胞のG0/G1期における細胞割合はG-CSF 0 ng/ml群に比べて有意に少なく（P < 0.05）、48時間および96時間培養後の正常CD34細胞のG0/G1期における細胞割合はG-CSF 0 ng/ml群に比べて有意に少なくなった。bcr/abl(+)-CD34+細胞および正常CD34+細胞のCD34の発現は培養時間とともに減少し、CD33およびCD13の発現は最初に増加し、後に減少したが、G-CSFの濃度とは相関しなかった。bcr/abl(+)-CD34+細胞および正常CD34+細胞は、増殖・分化とともに成熟した形態を示した。

RESULTS: The number of bcr/abl(+)-CD34+ cells was increased obviously in all groups. After cultured for 48 and 96 h, the number of bcr/abl(+)-CD34+ cells at G-CSF 10 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P < 0.05) , the number of normal CD34 cells was increased only in the presence of G-CSF. After cultured for 48, 96 and 144 h, the cell number in G-CSF 100 ng/ml group was significantly higher than that in G-CSF 0 ng/ml group (P < 0.05, P < 0.01, P < 0.01, respectively). After cultured for 144 h, the cell percentages in G0/G1 phase for bcr/abl(+)-CD34+ cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that in G-CSF 0 ng/ml group (P < 0. 05), and that for normal CD34 cells in G-CSF 10, 100, 1000 ng/ml groups were significantly less than that of G-CSF 0 ng/ml group after cultured for 48 and 96 h. The expressions of CD34 on bcr/abl(+)-CD34+ cells and normal CD34+ cells were decreased along with the culture duration, accompanied by the expression of CD33 and CD13 increased first and decreased later, which was not correlated with the concentration of G-CSF. Both bcr/abl(+)-CD34+ cells and normal CD34+ cells showed mature morphology along with proliferation and differentiation.

結論:

G-CSFはbcr/abl(+)-CD34+細胞と正常CD34+細胞の両方の増殖を促進するが、前者には不要であり、前者は後者よりも急速に分化するが、いずれもG-CSFとは無関係であった。

CONCLUSIONS: G-CSF promotes proliferation of both bcr/abl(+)-CD34+ cells and normal CD34+ cells, but not necessary for the former, and the former differentiates more rapidly than the latter does, but both was independent of G-CSF.