目的 研究半枝蓮總黃酮（TF-SB）對人胃腺癌細(xì)胞AGS增殖、凋亡和放療敏感性的影響和分子機(jī)制。方法 采用細(xì)胞計數(shù)試劑盒（CCK-8）法檢測TF-SB（0、25、50、100、200 μg·mL⁻¹）作用24 h對AGS細(xì)胞存活率的影響。將AGS細(xì)胞分為對照組、溶劑對照（含相同濃度的甲醇）組、TF-SB（100 μg·mL⁻¹）組、質(zhì)?？蛰d體（pcDNA，200 nmol·L⁻¹）組、遷移侵襲抑制蛋白（MIIP）過表達(dá)質(zhì)粒（pcDNA-MIIP，200 nmol·L⁻¹）組、TF-SB（100 μg·mL⁻¹）＋小干擾RNA對照（si-con，200 nmol·L⁻¹）組、TF-SB（100 μg·mL⁻¹）＋MIIP小干擾RNA（si-MIIP，200 nmol·L⁻¹）組，轉(zhuǎn)染質(zhì)粒后TF-SB處理24 h，CCK-8法檢測細(xì)胞存活率；流式細(xì)胞術(shù)檢測細(xì)胞凋亡；Western blotting法檢測B細(xì)胞淋巴瘤-xl（Bcl-xl）、裂解的半胱氨酸天冬氨酸蛋白酶（cleaved Caspase-3）、MIIP表達(dá)；不同照射量X射線（0、2、4、6、8 Gy）照射后，克隆形成實驗檢測細(xì)胞存活率，并繪制單擊多靶模型擬合曲線，Western blotting法檢測γ-H2AX蛋白表達(dá)。結(jié)果 與TF-SB 0 μg·mL⁻¹組比較，TFSB 25、50、100、200 μg·mL⁻¹組AGS細(xì)胞存活率顯著降低（P＜0.05）；與對照組及溶劑對照組比較，TF-SB 100 μg·mL⁻¹組AGS細(xì)胞凋亡率顯著升高（P＜0.05），Bcl-xl蛋白表達(dá)顯著降低（P＜ 0.05），cleaved Caspase-3和MIIP蛋白表達(dá)顯著升高（P＜0.05）；放療后，TF-SB組細(xì)胞存活率顯著降低（P＜0.05），γ-H2AX蛋白表達(dá)顯著升高（P＜0.05），放療敏感性增加。與pcDNA組比較，pcDNA-MIIP組AGS細(xì)胞凋亡率顯著增加，細(xì)胞存活率顯著降低，Bcl-xl蛋白表達(dá)顯著降低，cleavedCaspase-3蛋白表達(dá)增加（P＜0.05）；放療后，pcDNA-MIIP組細(xì)胞存活率顯著降低（P＜0.05），γ-H2AX蛋白表達(dá)顯著升高（P＜0.05），敏感性增加。與TF-SB＋si-con組比較，TF-SB＋si-MIIP組AGS細(xì)胞凋亡率顯著降低，細(xì)胞存活率顯著增加，Bcl-xl蛋白表達(dá)顯著增加，cleaved Caspase-3蛋白表達(dá)顯著降低（P＜0.05）；放療后，TF-SB＋si-MIIP組細(xì)胞存活率顯著升高（P＜0.05），γ-H2AX蛋白表達(dá)顯著降低（P＜0.05），敏感性降低。結(jié)論 TF-SB通過上調(diào)MIIP可抑制AGS細(xì)胞增殖，誘導(dǎo)細(xì)胞凋亡，提高其放療敏感性。

Objective To investigate the effect and molecular mechanism of total flavonoids of Scutellaria barbata (TF-SB) on the proliferation, apoptosis and radiosensitivity of gastric cancer cell AGS. Methods The cell counting kit (CCK-8) method was used to detect the effects of different concentrations (0, 25, 50, 100, 200 μg·mL⁻¹) of TF-SB on the survival of gastric cancer cells to determine the experimental concentration. AGS was divided into control group, solvent control group (methanol), TF-SB (100 μg·mL⁻¹), pcDNA (200 nmol·L⁻¹), pcDNA-MIIP (200 nmol·L⁻¹), TF-SB (100 μg·mL⁻¹) + si-con (200 nmol·L⁻¹), TF-SB (100 μg·mL⁻¹) + siMIIP (200 nmol·L⁻¹) group. Cell viability was detected by CCK-8 method. Flow cytometry was used to detect apoptosis. Clone formation test was used to detect the cell survival fraction after irradiation with different rays, and a one-click multi-target model was fitted to fit the curve. Western blotting was used to detect B cell lymphoma-xl (Bcl-xl), cleaved Caspase-3, migration invasion inhibitor protein (MIIP), γ -H2AX protein expression. Results Compared with TF-SB 0 μg·mL⁻¹ group, the survival rate of AGS cells in TF-SB 25, 50, 100 and 200 μg·mL⁻¹ groups was significantly decreased (P < 0.05). Compared with control group and solvent control group, the apoptosis rate of AGS cells in TF-SB 100 μg·mL⁻¹ group was significantly increased (P < 0.05), the expression of Bcl-XL protein was significantly decreased (P < 0.05), and the expression of Cleaved Caspase-3 and MIIP protein was significantly increased (P < 0.05). After radiotherapy, cell survival rate of TF-SB group was significantly decreased (P < 0.05), γ-H2AX protein expression was significantly increased (P < 0.05), and radiotherapy sensitivity was increased. Compared with the pcDNA group, the apoptosis rate of AGS cells in the pcDNA-MIIP group was significantly increased, the cell survival rate was significantly decreased, the expression of Bcl-XL protein was significantly decreased, and the expression of Cleaved Caspase-3 protein was increased (P < 0.05). After radiotherapy, the survival rate of pcDNA-MIIP group was significantly decreased (P < 0.05), the expression of γ-H2AX protein was significantly increased (P < 0.05), and the sensitivity was increased. Compared with TF-SB + si-con group, apoptosis rate of AGS cells in TF-SB + si-MIIP group was significantly decreased, cell survival rate was significantly increased, Bcl-XL protein expression was significantly increased, and Cleaved Caspase-3 protein expression was significantly decreased (P < 0.05). After radiotherapy, the survival rate of TF-SB + si-MIIP group was significantly increased (P < 0.05), the expression of γ -H2AX protein was significantly decreased (P < 0.05), and the sensitivity was decreased. Conclusions TF-SB could inhibit the proliferation of gastric cancer cell line AGS, induce apoptosis, and improve its radiotherapy sensitivity through up-reglating MIIP.

R285.5