目的 研究LZ-106對(duì)間變性淋巴瘤激酶（ALK）激酶活性的抑制作用，及對(duì)ALK陽(yáng)性細(xì)胞的生長(zhǎng)抑制作用。方法 采用分子對(duì)接模擬LZ-106與ALK的結(jié)合，通過(guò)體外激酶活性實(shí)驗(yàn)檢測(cè)其對(duì)ALK激酶活性的影響，通過(guò)CCK-8實(shí)驗(yàn)檢測(cè)其對(duì)H2228細(xì)胞的生長(zhǎng)抑制作用，通過(guò)Annexin V/PI細(xì)胞凋亡實(shí)驗(yàn)檢測(cè)其對(duì)H2228細(xì)胞的凋亡作用，通過(guò)Western blotting檢測(cè)其對(duì)H2228細(xì)胞p-ALK及下游信號(hào)通路關(guān)鍵激酶表達(dá)的影響。結(jié)果 LZ-106能抑制ALK激酶活性，半數(shù)抑制濃度（IC₅₀）為（15.51±2.09）nmol/L，其能顯著抑制H2228細(xì)胞存活率，并能濃度相關(guān)性地誘導(dǎo)細(xì)胞凋亡。Western blotting實(shí)驗(yàn)顯示LZ-106可以顯著抑制p-ALK、p-蛋白激酶B（Akt）及p-信號(hào)轉(zhuǎn)導(dǎo)因子和轉(zhuǎn)錄激活因子3（STAT3）的表達(dá)，且Akt及STAT3激活劑可以明顯削弱LZ-106的凋亡誘導(dǎo)作用。結(jié)論 LZ-106顯著抑制ALK激酶活性，誘導(dǎo)ALK陽(yáng)性細(xì)胞凋亡，激活A(yù)kt及STAT3可以逆轉(zhuǎn)LZ-106所致的細(xì)胞凋亡。

Objective To study the inhibitory effect of LZ-106 on ALK activity and growth inhibition of ALK positive cells. Methods The binding of LZ-106 to ALK was simulated by molecular docking, its effect on ALK kinase activity was detected by in vitro kinase activity assay, its growth inhibition effect on H2228 cells was detected by CCK-8 assay, and its apoptosis effect on H2228 cells was detected by Annexin V/PI apoptosis assay. Western blotting was used to detect its effects on the expression of p-ALK and key kinases in the downstream signaling pathway of H2228 cells. Results LZ-106 can inhibit the activity of ALK kinase with IC₅₀ of 15.51 ± 2.09 nmol/L, which can significantly inhibit the survival rate of H2228 cells and induce cell apoptosis in a concentration-dependent manner. Western blotting experiments showed that LZ-106 could significantly inhibit the expression of p-ALK, p-Akt, and p-STAT3, and Akt and STAT3 activators could significantly weaken the apoptosis-inducing effect of LZ-106. Conclusion LZ-106 significantly inhibits ALK kinase activity and induces apoptosis of ALK-positive cells. Activation of Akt and STAT3 can reverse the apoptosis caused by LZ-106.

R966

江蘇省自然科學(xué)基金資助項(xiàng)目（BK20151443），江蘇省中醫(yī)院院級(jí)課題（Y20038）