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本研究比较新型的酪氨酸激酶抑制剂HHGV678与伊马替尼(imatinib,IM)在体外对Bcr-Abl野生型和IM耐药细胞株的抑制作用,探索HHGV678替代IM治疗CML及IM耐药CML患者的可能性。以Bcr-Abl野生型细胞株(K562和32Dp210)及16种IM耐药细胞株(K562R和15种Bcr-Abl点突变细胞株)为研究对象,用MTT法检测HHGV678和IM对上述细胞的生长抑制作用;以DNA梯形条带法和Annexin-V/PI双染色流式细胞术检测细胞凋亡;应用Western blot检测HHGV678对上述细胞BCR-ABL融合蛋白及酪氨酸激酶磷酸化表达的影响。结果表明:HHGV678呈剂量依赖性显著抑制Bcr-Abl野生型细胞株和除T315I点突变细胞株以外的IM耐药细胞株生长。比较IC50发现,HHGV678在低剂量下(0.01-0.3μmol/L)抑制K562和32Dp210细胞生长的作用比IM分别强15.5和28倍;而对除T315I点突变细胞株以外的15种IM耐药细胞株细胞的生长抑制作用比IM强1.4-124.3倍。HHGV678抑制上述细胞酪氨酸激酶磷酸化的能力均强于IM。更重要的是HHGV678在10.0μmol/L剂量下诱导IM强耐药细胞株K562R和T315I点突变细胞株的凋亡率分别达到40.06%和33.32%,显著高于IM的19.77%和10.68%。结论:新型酪氨酸激酶抑制剂HHGV678对Bcr-Abl野生型细胞株和IM耐药细胞株,尤其是对IM强耐药细胞株的生长抑制作用明显强于IM,但HHGV678能否成为治疗CML和IM耐药CML患者新的靶向药物,仍有待进一步的研究。
This study compared the inhibitory effects of novel tyrosine kinase inhibitors HHGV678 and imatinib (IM) on Bcr-Abl wild-type and IM-resistant cell lines in vitro and explored the potential of HHGV678 for the treatment of CML and IM-resistant Possibility of CML patients. The Bcr-Abl wild-type cell lines (K562 and 32Dp210) and 16 kinds of IM resistant cell lines (K562R and 15 kinds of Bcr-Abl point mutant cell lines) were studied by MTT assay HHGV678 and IM on the growth of these cells The effect of HHGV678 on the phosphorylation of BCR-ABL fusion protein and tyrosine kinase was detected by Western blotting and DNA ladder method with Annexin-V / PI double staining. The results showed that HHGV678 significantly inhibited the growth of IM-resistant cell lines except Bcr-Abl wild-type cell line and T315I point-mutated cell line in a dose-dependent manner. Comparison of IC50 showed that HHGV678 inhibited the growth of K562 and 32Dp210 cells by 15.5 and 28 times respectively at low dose (0.01-0.3μmol / L), while 15 kinds of IM resistant cells except T315I point mutant cell line Growth inhibition of strain cells was 1.4-124.3 times stronger than IM. The ability of HHGV678 to inhibit the tyrosine kinase phosphorylation of these cells was stronger than that of IM. More importantly, the apoptotic rates of HHGV678 cells that were induced by 10.0μmol / L were 40.06% and 33.32% respectively, which were significantly higher than those of IM (19.77% and 10.68%). CONCLUSION: HHGV678 inhibits the growth of Bcr-Abl wild-type cells and IM-resistant cell lines, especially against IM-resistant cell lines significantly stronger than IM, but whether HHGV678 can be used as a treatment for CML And IM-resistant CML patients with new targeted drugs, remains to be further studied.