Application of small interfering RNA in preparing medicament for treating colon cancer
A small interference, colorectal cancer technology, applied in the field of biomedicine, can solve the problems of poor prognosis of patients with curative effect, and achieve the effect of inhibiting proliferation activity, retarding growth cycle, and inducing apoptosis of colorectal cancer cells
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Embodiment 1
[0027] Example 1 In vitro studies
[0028] The experimental materials of this example are human colorectal cancer cell line HCT116 purchased from the Institute of Cell Biochemistry, Chinese Academy of Sciences, and McCOY'S5A medium purchased from Sigma Company of the United States; SiRNA fragments targeting the four potential target sites of the mTOR gene and negative controls were synthesized by Dharmacon Company of the United States (Pre-designed RNAi Reagents, Dharmacon, USA); transfection reagent: DharmaFECT TM 4 siRNA Transfection Reagent (T-2004-03); 5×SiRNA Buffer: Dharmacon, USA; TRIzol Reagent: Invitrogen; MTT, Triton X-100, propidium iodide (PI): Sigma; RNase A: Roche; The sequences of siRNA, primers and probes used are shown in Table 1; the antibodies used in the test are shown in Table 2; kits for chemiluminescence detection: Super Signal West Femto Maximum Sensitivity Substrate(34095); Super Signal West Pico Chemilu-minescent Substrae (34077), both products o...
Embodiment 2
[0106] Example 2 In vivo studies
[0107] Experimental materials in this example Experimental animals: BALB / c nude mice, male, 4 weeks old, inbred strain, purchased from the Songjiang Animal Experiment Base of the Chinese Academy of Sciences (SCXK (Shanghai) 2003-0003), bred at the IVC of Shanghai Jiao Tong University School of Medicine Animal laboratory (SYXK (Shanghai) 2003-0007); siRNA: using the siRNA3 and negative control (NegativeControl) sequences shown in Table 1, modified with methoxy by Shanghai GenePharma Co., Ltd, and chemically synthesized ; Transfection reagent: in vivo-jetPEI TM (201-50G, Polyplus Transfection, Illkirch, France); Antibodies used in the test are shown in Table 2; TUNEL Apoptosis Detection Kit: Roche, 1684809a; 50 μl micro-syringe: Shanghai Tiancheng Technology Co., Ltd.; Vernier caliper: Shanghai Precision Instruments Co., Ltd.; ultrasonic cell pulverizer (JY92-II): Ningbo Xinzhi Biotechnology Co., Ltd.; paraffin slicer (RM21459), embedding mach...
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