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肺がん幹細胞の単離と生物学的特性評価
[Isolation and Biological Characterization of Lung Cancer Stem Cells].
PMID: 32131936 DOI: 10.3881/j.issn.1000-503X.10911.
抄録
肺がん幹細胞のスクリーニング方法と生物学的特徴を探る。 ADMで誘導したSPC-A-1/アドリアマイシン(ADM)細胞株からABCG2細胞とABCG2細胞を選抜し、ヌードマウスに移植してフローサイトメトリーで腫瘍性を解析した。 その結果、SPC-A-1細胞の平均蛍光強度は(1.001±0.014)×10であり、SPC-A-1/ADM細胞の蛍光強度(10.257±0.023)×10に比べて有意に低い値であった(=17.320,=0.001)。320,=0.001）；また、ABCG2/BCRP-FITC処理群とSPC-A-1対照群との間にも統計的に有意な差が認められた（=5.269,=0.021）；SPC-A-1対照群との間にも統計的に有意な差が認められた（=6.869, =0.012）、SPC-A-1/ADM細胞対照群とSPC-A-1/ADM細胞ホモタイプ対照群（=8.112,=0.015）の間で、ABCG2/BCRP-FITCで処理したSPC-A-1/ADM細胞の陽性率はSPC-A-1細胞の9.8％、39.84倍であり、ABCG2/BCRP-FITC群とSPC-A-1/ADM群（=9.120,=0.005）との間で有意な差が認められた。B群の陽性率はA群の684倍であり、その差は統計的に有意であり(=11.235,=0.001)、B群の蛍光強度も強く、SPC-A-1細胞、A群、B群を接種したマウスの平均腫瘍化体積は(6.96;1.82)であった。B群の腫瘍化率は75.00%であり、SPC-A-1細胞やA群に比べて有意に高かった(=19.780,=0.002)。 ヒト肺腺癌SPC-A-1/ADM細胞株からABCG2抗体と免疫磁気ビーズソーティング法を組み合わせてABCG2細胞を分離し、ヌードマウスでの腫瘍形成率を観察することで、肺癌幹細胞の同定と生物学的特徴を探ることができる。
To explore the methods of screening and biological characteristics of lung cancer stem cells. We selected the ABCG2 and ABCG2 cells from SPC-A-1/adriamycin(ADM)cell line induced by ADM and analyzed the tumorigenicity of ABCG2 and ABCG2 cells by flow cytometry and transplantation in nude mice. The average fluorescence intensity of SPC-A-1 cells was(1.001±0.014)×10 ,which was significantly lower than that of SPC-A-1/ADM cells [(10.257±0.023) ×10 ](=17.320,=0.001);the difference was also statistically significant between the ABCG2/BCRP-FITC treatment group and the SPC-A-1 control group(=5.269,=0.021) and the SPC-A-1 control group(=6.869, =0.012) and between the SPC-A-1/ADM cell control group and the SPC-A-1/ADM cell homotype control group(=8.112,=0.015).The positive rate of SPC-A-1/ADM cells treated with ABCG2/BCRP-FITC was 9.8%,39.84 times higher than that of SPC-A-1 cells;it showed significant difference between the ABCG2/BCRP-FITC group and the SPC-A-1/ADM group(=9.120,=0.005) and the SPC-A-1/ADM group(=8.257,= 0.006).The positive rate of group B cells was 684 times that of group A cells,and the difference was statistically significant(=11.235,=0.001),and the fluorescence intensity of group B cells was strong.The average tumorigenic volume of the mice inoculated with SPC-A-1 cells,group A cells,and group B cells was(6.96±1.82),(6.70±2.55),and(9.17±2.41) mm ,respectively.Among them,group B was the highest,but there was no significant difference among these three groups(=2.362,=0.086).The tumorigenic rate of group B cells was 75.00%,which was significantly higher than that of SPC-A-1 cells and group A cells(=19.780,=0.002). ABCG2 cells from human lung adenocarcinoma SPC-A-1/ADM cell line can be isolated by ABCG2 antibody combined with immunomagnetic beads sorting method,and the tumor formation rate in nude mice can be observed to explore the identification and biological characterization of lung cancer stem cells.