Mir 30a 5p agomir

PDAC-bearing nude mice with subcutaneous passage of PANC-1-luciferin were used. When the tumor size reached approximately 5 mm in length, the mice were divided into two groups (four mice per group) randomly. Stabilized microRNAs (miRNAs) (miR-30a-5p agomir and negative control agomir) were purchased from RiboBio (miR40000087-1-10; Guangzhou, China). miR-30a-5p agomir (5 nM) or control oligo mixture was injected into the xenografts in a multi-site injection manner every 3 days for 2 weeks. The tumor volume was measured with a caliper every 3 days, using the following formula: volume = length × width²/2. At the end of the 30-day observation period, the mice bearing tumor xenografts were examined by bioluminescence imaging and then sacrificed and the tumor tissues were removed for formalin fixation.
The animals anesthetized by isoflurane were intraperitoneally injected with d-luciferin (Biotium, USA) in a concentration of 150 mg/kg and, 20 min later, were subjected to the in vivo bioluminescence imaging using the system of photobiology.

PDAC-bearing male nude mice with subcutaneous passage of BxPC-3 were used. When the tumor size reached approximately 5 mm in length, the mice were divided into two groups (3 mice per group) randomly. Stabilized miRNAs (miR-30a-5p agomir and Negative control agomir) were purchased from RiboBio (miR40000087-1-10, Guangzhou, China). miR-30a-5p agomir (5 nM) or control oligos mixture was injected into the xenografts in a multi-site injection manner every 3 days for two weeks, and then followed by administration with gemcitabine (100 mg/kg). At the end of the 30-day observation period, mice were killed and the tumors were measured. The tumor volume was measured with a caliper, using the formula volume = length × width2 (link)/2.

Female BALB/c athymic nude mice (4–6 weeks old and weighing 16–20 g) were purchased from the Experimental Animal Center of Soochow University and bred under pathogen-free conditions. All the animal experiments were carried out in accordance with the Guide for the Care and Use of Experimental Animals Center of Soochow University. To establish the lung carcinoma xenograft model, A549 cells and stable CD73-knockdown A549 cells were suspended in 100 ml PBS and inoculated subcutaneously into the flanks of nude mice. After 8–10 days, the nude mice with transplanted A549 cells were randomly divided into two groups (8 mice in each group). The mice with transplanted CD73-knockdown A549 cells were also split into two groups. An miR-30a-5p agomir and NC agomir (RiboBio Co. Ltd., Guangzhou, China) were directly injected into the A549 implanted tumors at a dose of 2 nmol (in 20 μl PBS) per mouse every 4 days seven times (28 days total). Chemically stabilized miRNAs may have markedly improved pharmacological properties, as described before [44 (link)]. Tumor volume (V) was determined by measuring the length (L) and width (W) with a vernier caliper and applying the formula V = (L × W²) × 0.5.