动物造模,肿瘤模型 z-bio 2021-04-08 406

　　There are many ways to replicate animal tumors. For example, laboratory animals can be irradiated, radioisotopes can be injected intravenously or locally, and various chemical carcinogens (alkylating agents, polycyclic aromatic hydrocarbons, aromatic amines, amino groups can be used). Azo dyes, etc.), the use of ammonium nitrate) or the use of plant toxins (psychados, saflors, etc.); the use of metals (chromium, nickel, arsenic, cadmium, etc.); the use of RNA and DNA cancer viruses; the use of various carcinogenic fungal toxins (most (Carcinogenic) They are aflatoxins and can induce a variety of tumors. The number of induced tumor models is the first among induced animal models. Carcinogens are commonly used to cause tumors in animals through oral administration, injection, implantation, and smears.

　　There are many reports that viruses can induce tumors in animals. For example, murine leukemia virus (MLV), avian leukemia virus (ALV), and feline leukemia virus (FLV) can cause leukemia in rats, mice, chickens, and cats. ,respectively. The loose chicken cancer virus can cause sarcoma in a variety of animals, including voles, chickens, ducks, quails, monkeys, and snakes. Feline Sarcoma Sarcoma (FSV) can cause sarcoma in rats, cats, dogs and monkeys. Human adenovirus can induce sarcoma and lymphoma in mice and field mice.

　　(1) Induced tumor animal model

　　1. Liver cancer Diethylnitrosamine (DEN) induces liver cancer in rats: a group of male and female rats are sealed, weighing about 250 g. They are kept in cages by gender. In addition to regular food, it also provides carcinogens. That is, a 0.25% DEN aqueous solution is used to force the stomach to be fed at a dose of 10 mg/kg once a week, and the 0.025% DEN aqueous solution is put into a water bottle. You can drink for free for the remaining 5 days. In total, it can induce liver cancer in about 4 months. Or, mix 0.005% of water and drinking water orally for 8 months and use a single dose to induce liver cancer. 4-2 Methylaminobenzene (DBA) induces liver cancer in rats: The rats are fed a 0.06% DBA diet, and the vitamin B2 in the diet should not exceed 1.5-2 mg/kg. 4 Induce a large number of liver cancers. -6 months. success. 2-Acetyl amino acid (2AAF) induces liver cancer in mice, dogs, cats, chickens and rabbits. Adult rats were fed a standard diet containing 0.03% 2AAF. An average of 2-3 mg of 2AAF per day (2AAF can also be mixed with oil and fed), after 3-4 months, 80-90% of animals will develop liver tumors. Diethylnitrosamine induces liver cancer in rats. The dose was 0.3-14 mg/kg body weight/day, mixed with feed or drinking water, and 255/300 rats developed liver cancer after 6-9 months. Aminoazotoluene (OAAT) induces liver cancer in mice: Put 1% OAAF benzene solution (about 0.1 ml containing 1 mg) on the skin between the shoulders of the animal once every other day, 2-3 drops each time, usually 100 drops each time . The first liver tumor appeared 7-8 weeks after the experiment, which can induce about 55% of mouse liver tumors within 7 months or longer. Alternatively, using 2.5 mg of OAAT dissolved in sunflower seed oil, subcutaneously injected into C3H mice four times a day at 10 days intervals may induce liver cancer. Aflatoxin can induce liver cancer in rats: the daily diet contains 0.001 to 0.015 ppm. After mixing with feed for 6 months, the induction rate of liver cancer reached 80%.

　　2. Gastric cancer methylanthraphenol induces gastric cancer in mice: Take about 20 g of mice and pass through the methylcholanthracene (MC) knot on the surface of the glandular gastric mucosa under aseptic surgery. The knot of the MC-containing wire is made of ordinary thin wire. After tying the ligature to one end, place the ligation in a small glass test tube containing MC, and then slightly heat the alcohol lamp to liquefy and infiltrate the MC. Knot. Put 20-methylcholestene with an MC concentration of 0.05 to 0.1 g in 10 to 20 threads. Gastric cancer can be successful within 4-8 months after surgical implantation. In the case of an asymmetric nitrosamine dose of 0.25 ml/kg body weight, all animals will develop anterior gastric papillary carcinoma after 3 months, and 85-100% of animals will develop pregastric carcinoma after 7-8 months. Stomach cancer. Kunming is the most sensitive. Line A is second, and line 615 is the least sensitive. In addition, urea methyl nitrosoacetate can be used to add 2 mg/kg body weight to the drinking water of BD rats 5 times a week. After 520 days, all rats developed glandular gastric cancer.

　　3. Esophageal cancer methylbenzylnitrosamine (MBNA) induces esophageal cancer in rats. Ingest Wistar rats weighing more than 100 grams, feed them with drinking water containing methylbenzylnitrosamine, and mix MBNA with the diet, with a daily intake of 0.75 to 1.5 mg/kg body weight. 80-100 days can induce esophageal cancer. You can also use dihydrosafrole, which is a flavoring for beer. Adding 2,500 to 10,000 ppm of safrole in the diet of rats can cause 20% to 75% of esophageal cancers. A 0.2% or 0.005% methylbenzylnitrosamine aqueous solution was used orally to give animals. Rats were perfused at a dose of 1 mg/kg bw once a day, and a case of esophageal papilloma was found on day 27 and day 154. The first case of esophageal cancer was 53% at 11 months.

　　Four. Lung cancer Diethylnitrosamine (DEN) induces lung cancer in mice. The mice were injected subcutaneously with 1% DEN aqueous solution once a week, each dose reached 56 mg/kg, the total dose of DEN reached 868 mg, and the observation time was about 100 days. .. The pass rate can reach 40%. When the total dose of DEN reaches 1176 mg, the observation time is about half a year, and the incidence of cancer can reach 94%. Carbamate-induced lung adenocarcinoma: Mice (line A, 1-11/2 months old) are more sensitive than rats, and 0.1-0.3 ml of 10% carbamate is injected intraperitoneally every 3-5 days Ester saline. Note: A total of 2-3 months of injection, the dose of each mouse is about 100 mg, the incidence of lung adenocarcinoma 3 months after injection is 100%, most of which are multiple. This type of induced tumor is benign. In addition, intratracheal injections such as benzopy, ammonium sulfate aerosol and methylanthracene can also be used to induce lung cancer. For example, 3,4 benzore was injected into the trachea of monkeys (3-15 mg of benzoic acid and the same amount of Fe2O3 mixture) once a week for a total of 10 injections. 2 out of 6 monkeys had lung squamous cell carcinoma Induced in animals. Someone inhaled amine sulfate aerosol in 100 rats. Thirteen months later, all rats developed lung adenocarcinoma. The methoxanthene was mixed with 0.2% gelatin to form a suspension and injected into the trachea of a golden hamster. 0.1 ml (containing 5 mg of methylcholine) once a week for a total of 6 times. After 53 weeks, the animal became ill. Lung cancer.

　　Fives. Nasopharyngeal carcinoma Dimethylcholantrene (MC) induces nasopharyngeal carcinoma in rats: Take a hard plastic rice tube with a diameter of 2-3 mm and pull it into the cone of an alcohol lamp with a small flame. Each part is about 3.5 cm long and contains a crystal MC equipped with a tube. One end of the small tube was fire-sealed to prevent the drug from spilling, and the tip was pierced with a needle to allow MC to overflow the penis. Both males and females need to take about 120 grams of white mice. After ether anesthesia, insert the above-mentioned plastic tube containing MC into the nasal cavity through the front nostril. It is characterized by small and thick front nostrils. Application. Push a small tube into all objects with very little force, and the tip can reach the nasopharyngeal cavity. Without additional fixation, the small tube can stay in the nasal cavity for a long time. If the animal dies naturally within a given time (more than half a year), the nasopharynx is fixed with 10% formalin, decalcified, embedded in paraffin, and made into serial sections. The incidence of cancer can reach 60% or higher. Nasopharyngeal carcinoma induced by diethylnitrosamine rhinorrhea: Take about 120 g of hermaphrodite white mice, anesthetize ether, and then use a flat pointed 8 gauge needle to gently insert the anterior nostril, and the needle tip reaches the nostril cavity. . Newly prepared 0.02 ml (containing DEN 6.7 mg) 33.3% DEN suspension of 1% Tween-80, injected 15-20 times a week, may induce nasopharyngeal cancer.

　　6. Take a female mouse with cervical cancer and use a vaginal dilator and a sharp curved needle to tie the thread to the cervix under the state of an unanaesthetized animal. Pass behind the right uterine horn and fix the cervical knot. The other end of the thread is fixed to the muscles of the back and the skin is sutured. After hanging up the line, I injected penicillin for 2-3 days on the same day. Prevent postoperative infection. At a certain time point (about half a year), the animals were sacrificed, and the cervical tissues were fixed with 10% formalin, embedded in paraffin and serially sectioned.

　　7. Colon cancer: Four-week-old male rats were injected subcutaneously with dimethylhydrazine (DMH) for 21 consecutive weeks at 21 mg/kg DMH each time. The animals were sacrificed 1-4 weeks after the last dose. The descending colon was fixed with Buan solution, dehydrated, embedded in paraffin and cut into thin slices. First, prepare 400 mg of DMH per 100 ml of mother liquor, then add 37 mg of EDTA, and adjust the pH to 6.5 with sodium hydroxide (0.1N) solution for later use. (2) Transplantable animal model of tumor

　　Most of the anti-tumor drugs currently used in clinical practice are discovered through screening in animal transplantation tumor trials. The benefits of using animal xenograft screening drugs are: a group of animals are simultaneously inoculated with the same number of tumor cells, the growth rate is relatively consistent, individual differences are small, and the survival rate of the inoculation is almost 100%. , The effect on the host is similar, it can be continuously transplanted into animals of the same species or strain, so that its therapeutic effect can be judged objectively and easily, and can be used for long-term research. The test cycle is usually very short, but the test conditions are also easy to control. Therefore, most current anti-tumor screening uses animal xenografts as the screening model. Currently, there are about 400 animal transplanted tumors stored in the world, but usually only 20 to 30 tumors are used in screening tests. According to statistics in 1984, my country has established 64 common tumor strains of different animals and humans in the same strains and types of animals. For example, mouse lung adenoma (HP615), mouse neck tumor number 27 (U27), mouse brain tumor number 22 (B22), mouse lymphopenia (L615), nude mouse human liver tumor xenografts and Human brain malignant glioma (NCS-1)) and so on. Animal tumors can be transplanted and subcultured to cultivate the desired tumor cell lines. Tumor strains are tumor cell models with stable histology and proliferation characteristics, and can be serially passaged in the same strain or animal. Transplanting tumors into healthy animals is equivalent to tissue culture, and the types of tumors can be preserved for a long time for experiments. In experiments, ascites and solid tumors are usually used for transplantation. For tumors that produce ascites, a specific number of cells can be injected into the abdominal cavity of the recipient animal to form ascites tumors or produce ascites. The transplantation of solid tumors can also be performed aseptically. The solid tumors are cut into small pieces of 2-3 mm and then transplanted under the skin of the recipient animal.

　　Tumor transplantation from autologous animals does not cause excretion. The transplantation of the same animal can be injected with corticosteroids, anti-tumor drugs and appropriate amount of radiation to reduce the host's immune rejection. Xenogeneic tumor transplantation began in Leidy (1834), but it was more difficult. In the past 50 years, the following methods have been commonly used in allogeneic transplantation: (1) Inoculation under the skin or submucosa has the advantage of being easy to observe, but the rejection is large and the effect is insufficient. (2) Transplanting animal tumors into chicken embryo allantoic. One feature is that it is easy to survive, but human tumors have not been well reported. ③Inoculate human tumors into the anterior chamber of rats, guinea pigs and rabbits. The disadvantage is that the cells cannot be passaged. ④ Transplant into the brain of an animal. The tumor grows rapidly, but it is difficult to observe. In 1983, Badgen et al. used athymic rats to transplant human tumors under the kidney capsule to screen for new drugs. All experiments took only 11 days, and the hit rate was very high. This study provides good news for drug screening in clinical patients. Since the 1960s, hundreds of transplantable human tumors have been established abroad. These tumor strains can prevent the formation and function decline associated with passage. In 1969, Rygaaid took the lead in successfully transplanting human tumors into athymic nude mice, opening up a new field for allogeneic tumor transplantation. The lack of T lymphocyte function in nude mice makes it a highly ideal tumor transplantation model material.