Gemcitabine hydrochloride (LY 188011 hydrochloride) is a synthetic cytosine nucleoside derivative and an inhibitor of DNA synthesis. Gemcitabine has antitumor and antimetabolic activities. Gemcitabine induces autophagy and apoptosis.

DNA synthesis (Mia Paca-2 cells):40nM, DNA synthesis (BxPC-3 cells):18nM, DNA synthesis (PANC-1 cells):92.7nM, DNA synthesis (PL-45 cells):89.3nM, DNA synthesis (Capan2 cells):12nM

方法：PDAC 衍生成对原发性癌症细胞 (PCCs) PCC-1、PCC-2、PCC-5、PCC-6 和 PDAC 细胞 BxPC-3、 Mia PaCa-2、Panc-1 用 Gemcitabine hydrochloride (0.001-1000 µM) 处理 48 h，使用 MTT 方法检测细胞生长抑制情况。
结果：Gemcitabine 剂量依赖性地抑制 PCC-1、PCC-2、PCC-5、PCC-6、BxPC-3、 Mia PaCa-2、Panc-1 细胞生长，IC50 分别为 1.2/0.3/1.2/4.3/4.2/7.9/10.5 µM。[1]
方法：人胰腺癌细胞 PK-1 用 Gemcitabine hydrochloride (30 nM) 处理 24-48 h，使用 Flow Cytometry 方法检测细胞周期情况。
结果：Gemcitabine 诱导 PK-1 细胞在 G0/G1 期的百分比增加，S 期及 G2/M 细胞比例下降。Gemcitabine 诱导 PK-1 细胞 S 期细胞周期停滞。[2]
方法：人肺癌细胞 SPC-A1 和 A549 用 GFP 标记的 LC3 转染，24 h 后 加 Gemcitabine hydrochloride (5 μM) 孵育 24 h，使用共聚焦激光扫描显微镜检测 LC3 表达情况。
结果：LC3-II 的积累是自噬的标志。 Gemcitabine 显著增加了肿瘤细胞的 GFP-LC3 点，表明自噬水平提高。[3]

方法：为检测体内抗肿瘤活性，将 Gemcitabine hydrochloride (20 mg/kg) 腹腔注射给携带人高级别脑膜瘤肿瘤 HKBMM 的 BALB/cAJcl-nu/nu 小鼠，每周两次，持续四周。
结果：Gemcitabine 治疗不仅抑制了肿瘤的发生，而且抑制了肿瘤生长。Gemcitabine 在体内阻断肿瘤细胞的细胞周期进程并促进细胞凋亡。Gemcitabine 通过细胞抑制和细胞毒性机制对高级别脑膜瘤发挥强大的抗肿瘤活性。[4]
方法：为检测体内抗肿瘤活性，将 Gemcitabine hydrochloride (50 mg/kg/每周两次/腹腔注射) 和 DMAPT (40 mg/kg/天/灌胃) 给携带胰腺癌肿瘤的 LSL-KrasG12D/+; LSL-Trp53R172H; Pdx-1-Cre 突变小鼠。
结果：Gemcitabine 或 DMAPT/Gemcitabine 组合显著提高了中位生存率 (254.5 或 255  天与 217.5 天)，并降低了胰腺腺癌的发生率和多样性。Gemcitabine 治疗增加了小鼠血浆中 IL-1α、IL-1β和IL-17的水平，而 DMAPT/Gemcitabine 降低了 IL-12p40、MCP-1、MIP-1β、eotaxin 和 TNF-α的水平，都是 κB 的靶基因。[5]

The cytotoxic effect of gemcitabine was evaluated with the MTT assay. SPC-A1 or A549 cells were treated with gemcitabine (0.05–500 lM) for 24 h. Then, 10 ll of MTT (5 mg/ml in PBS) was added to each well and incubated for 4 h at 37 C. Then, the formazan crystals were solubilized with 200 ll DMSO. The absorbance at 570 nm was measured using an automatic multiwell spectrophotometer. The experiment was repeated four times for each group [3].

At 1 month of age, LSL-Kras G12D/+; LSL-Trp53 R172H; Pdx-1-Cre mice are randomized into treatment groups (placebo, DMAPT, Gemcitabine, DMAPT/Gemcitabine). Placebo (vehicle=hydroxylpropyl methylcellulose, 0.2% Tween 80 [HPMT]) and DMAPT (40 mg/kg body weight in HPMT) are administered by oral gastric lavage once daily. Gemcitabine (50 mg/kg body weight in PBS) is administered by intraperitoneal injection twice weekly. Mouse weight is monitored weekly. Treatment is continued until mice show signs of lethargy, abdominal distension or weight loss at which time they are sacrificed. Successful excision-recombination events are confirmed in the pancreata of mice by detecting the presence of a single LoxP site [5].