目的 研究金復(fù)康口服液對人肺腺癌PC9細(xì)胞及耐藥PC9/R細(xì)胞的吉非替尼增敏作用及機(jī)制。方法 金復(fù)康口服液20 mg/mL聯(lián)合不同濃度的吉非替尼（40、20、10、5、2 μmol/L）作用于PC9/R細(xì)胞，CCK-8法檢測細(xì)胞增殖；培養(yǎng)PC9、PC9/R細(xì)胞，分為對照組、金復(fù)康口服液組、吉非替尼組、聯(lián)合用藥組，流式細(xì)胞術(shù)檢測PC9、PC9/R細(xì)胞周期和凋亡。在BALB/c裸鼠右前肢腋窩皮下接種PC9/R細(xì)胞建立裸鼠移植瘤模型，觀察金復(fù)康口服液聯(lián)合吉非替尼的體內(nèi)抑瘤作用；對AKT、p-AKT、PTEN、PDCD4蛋白表達(dá)的影響和對miRNA-21表達(dá)的影響。結(jié)果 與吉非替尼（2、5、10、20 μmol/L）組比較，吉非替尼（2、5、10、20 μmol/L）+金復(fù)康口服液（20 mg/mL）組的PC9/R細(xì)胞抑制率顯著增加（P<0.01）；與吉非替尼組比較，聯(lián)合用藥可以通過顯著增加早期凋亡細(xì)胞比例誘導(dǎo)PC9及PC9/R細(xì)胞凋亡，并使PC9、PC9/R細(xì)胞停滯在DNA合成前期，從而抑制細(xì)胞增殖，差異均具有統(tǒng)計學(xué)意義。體內(nèi)實驗表明，與吉非替尼組比較，聯(lián)合用藥顯著抑制裸鼠PC9/R移植瘤的生長（P<0.05），且能顯著降低裸鼠PC9/R組織p-AKT表達(dá)（P<0.05），增強(qiáng)PTEN、PDCD4的表達(dá)（P<0.05），聯(lián)合用藥組裸鼠PC9/R瘤組織miRNA-21的表達(dá)顯著降低（P<0.05）。結(jié)論 金復(fù)康口服液能提高人肺腺癌PC9/R細(xì)胞對吉非替尼的敏感性，其作用的可能機(jī)制為通過降低miRNA-21的表達(dá)從而增強(qiáng)PTEN、PDCD4的表達(dá)以抑制AKT通路活性。

Objective To study investigate the mechanism of Jinfukang Oral Liquid in increasing the sensitivity of human lung adenocarcinoma PC9/R cells to gefitinib. Methods The proliferation of PC9/R cells was detected by CCK-8 method, after Jinfukang Oral Liquid 20 mg/mL combined with different concentrations of gefitinib (40, 20, 10, 5, 2 μmol/L) treating on PC9/R cells. Flow cytometry was used to detect the effects of each group on cell cycle and apoptosis of PC9 and PC9/R cells. PC9/R nude mice model was established by subcutaneous inoculation of PC9/R cells into the axilla of right forelimb of BALB/c nude mice. To observe the effect of Jinfukang oral liquid on the anti-tumor activity of gefitinib, the expression of AKT, p-AKT, PTEN and PDCD4 and the expression of microRNA-21 in nude mice xenografted with PC9/R cells. Results Compared with gefitinib (2, 5, 10, 20 μmol/L) group, the inhibition rate of PC9/R cells in gefitinib (2, 5, 10, 20 μmol/L) + Jinfukang Oral Liquid (20 mg/mL) group increased significantly (P<0.01). The combination of Jinfukang oral liquid and gefitinib could induce PC9 and PC9/R cell apoptosis by increasing the proportion of early apoptotic cells and arrest PC9 and PC9/R cells in the early stage of DNA synthesis, thus inhibiting cell proliferation. In vivo, the combination of PC9/R could inhibit the growth of PC9/R xenografts in nude mice more effectively and significantly by reducing the expression of pAKT in PC9/R tissues of nude mice (P<0.05), enhancing the expression of PTEN and PDCD4 (P<0.05). Notably, PC9/R inhibited the expression of microRNA-21 in the tumor tissues of nude mice (P<0.05). Conclusion Jinfukang Oral Liquid can enhance the sensitivity of human lung adenocarcinoma PC9/R cells to gefitinib, and its possible mechanism is to enhance the expression of PTEN and PDCD4 by reducing the expression of microRNA-21, thus inhibiting the activity ofAKT pathway.

R285.5

國家自然科學(xué)基金資助項目（81573890）；國家中醫(yī)藥管理局中醫(yī)臨床研究基地業(yè)務(wù)建設(shè)科研專題課題（JDZX2015069）；劉嘉湘國醫(yī)大師傳承工作室建設(shè)項目（DSGZS-2017002）；龍華醫(yī)院科技創(chuàng)新項目（KY2017、KY1739）；上海中醫(yī)藥大學(xué)附屬龍華醫(yī)院中青年名中醫(yī)培養(yǎng)項目