目的 研究尼美舒利聯(lián)合吉西他濱對(duì)人非小細(xì)胞肺癌NCI-H1975細(xì)胞增殖和凋亡的影響。方法 采用MTT法測(cè)定尼美舒利聯(lián)合吉西他濱對(duì)NCI-H1975細(xì)胞生長(zhǎng)的抑制作用。流式細(xì)胞儀檢測(cè)尼美舒利（100 μmol/L）組、吉西他濱（20 nmol/L）組和聯(lián)合用藥（尼美舒利100 μmol/L+吉西他濱20 nmol/L）組對(duì)NCI-H1975細(xì)胞周期和凋亡的影響。Western blotting檢測(cè)各組對(duì)Bcl-2、Bax、Cleaved caspase-3蛋白表達(dá)的影響。結(jié)果 尼美舒利對(duì)NCI-H1975細(xì)胞的生長(zhǎng)具有抑制作用，且呈劑量相關(guān)性，IC₅₀為（298.76±2.79）μmol/L；吉西他濱的IC₅₀為（78.64±3.14）nmol/L，尼美舒利與吉西他濱聯(lián)合作用后，能明顯增加后者對(duì)NCI-H1975細(xì)胞增殖的抑制作用，二者有明顯的協(xié)同作用。尼美舒利聯(lián)合吉西他濱將細(xì)胞阻滯在G₀/G₁期，而S和G₂/M期均有所減少。與單藥組相比，聯(lián)合用藥組能顯著升高細(xì)胞的凋亡率（P<0.05）。與單藥組相比，聯(lián)合用藥組明顯下調(diào)Bcl-2蛋白的表達(dá)（P<0.05），上調(diào)Bax和Cleaved caspase-3蛋白的表達(dá)（P<0.05）。結(jié)論 尼美舒利聯(lián)合吉西他濱能明顯抑制腫瘤細(xì)胞的增殖，誘導(dǎo)細(xì)胞凋亡，其效果優(yōu)于單用吉西他濱組，其機(jī)制與影響細(xì)胞周期調(diào)控，下調(diào)Bcl-2蛋白表達(dá)、上調(diào)Bax和Cleaved caspase-3蛋白表達(dá)水平有關(guān)。

Objective To study the effects of nimesulide combined with gemcitabine on proliferation and apoptosis of human non-small cell lung cancer NCI-H1975 cells. Methods MTT assay was used to determine the inhibitory effect of nimesulide combined with gemcitabine on the growth of NCI-H1975 cells. Flow cytometry was used to detect the cell cycle and apoptosis of NCI-H1975 in the nimesulide (100 μmol/L) group, the gemcitabine (20 nmol/L) group, and the combination (nimesulide 100 μmol/L + gemcitabine 20 nmol/L) group. The expression of Bcl-2, Bax, and Cleaved caspase-3 protein was detected by Western blotting. Results Nimesulide inhibited the growth of NCI-H1975 cells in a dose-dependent manner with an IC₅₀ of (298.76 ±2.79) μmol/L. The IC₅₀ of gemcitabine was (78.64 ±3.14) nmol/L. The combination of nimesulide and gemcitabine significantly increased the inhibitory effect of the latter on the proliferation of NCI-H1975 cells. The ratio of nimesulide plus gemcitabine to cells in the gemcitabine group was significantly increased in the G₀/G₁ phase, while the S and G₂/M phases were reduced. Compared with the single-agent group, the apoptosis rate in the combined group was significantly increased (P < 0.05). Compared with the single-agent group, the combination group could significantly down-regulate Bcl-2 protein expression (P < 0.05), but up-regulate Bax and Cleaved caspase-3 protein expression (P < 0.05). Conclusion Nimesulide combined with gemcitabine can significantly inhibit the proliferation of tumor cells and induce apoptosis. The effect is better than that of gemcitabine alone. The mechanism and effect of cell cycle regulation, down-regulation of Bcl-2 protein expression, up-regulation of Bax and Caspase-3 protein expression levels are related.

R966

國(guó)家自然科學(xué)基金資助項(xiàng)目（81472474）；天津市衛(wèi)生局科技基金資助項(xiàng)目（201242034）