Orthotopic nude mice model of HCC and treatment Male athymic BALB/c nu/nu mice, 4 six weeks old, were obtained from Beijing HFK Bio Technologies Co. Ltd and housed in particular pathogen no cost ailment with the Animal Experiment Center of Wuhan University. All animal experiments have been carried out in accordance with the suggestions and accepted protocols of your University of Wuhan Animal Experiment Center Institutional Animal Care and Use Committee. Nude mice model with spontaneous pulmonary metas tasis was established as described previously. Briefly, HCCLM9 cells in 0. 1 ml phosphate buffered saline had been injected subcutaneously in to the left upper flank of two nude mice. The subcutaneous tumors have been eliminated whenever they reached 8 mm in diameter, and minced into pieces to execute orthotopic transplantation into livers of nude mice.
On day 8 after model establishment, the mice have been randomized into Management group, DOX group, and PDOX group, and treated as illus trated from the movement chart of Figure 1. In our previous review, we found that by intraperi toneal injection, PDOX could possibly be securely administered at twice the dose of DOX on a molar basis. Consequently, we improved the doses of the two DOX and PDOX read the article within this review, by which the PDOX dose was three times that of DOX on the molar basis. The behaviors and standard disorders were monitored regular, and body weights of animals were recorded twice a week. On d 36 publish inoculation, the mice were anes thetized by peritoneal injection of 3% phenobarbital chloride, and after that subjected to magnetic resonance picture study to measure the liver tumor size.
The peripheral blood was obtained for schedule blood tests and biochemis look at scientific studies. Tumor tissues and main organs which includes the heart, the liver, the lungs and any other suspected organs were collected, fixed with 4% paraformaldehyde Fosbretabulin disodium and embedded in paraffin for pathological and immuno histochemistry research. Also, fresh tumor tissues had been obtained for western blotting studies as thorough below. Experimental peritoneal carcinomatosis index An experimental peritoneal carcinomatosis index strategy was developed to assess the efficacy that took into consideration of tumor nodule sizes, distributions, as well as characteristics of ascites. Within this strategy, the stomach cavity from the mouse was divided into four regions, area I, subdiaphragm, region II, the liver, spleen, stom ach, and affiliated ligaments, area III, the modest intestine, colon, mesenterium, and abdominal wall, and region IV, the pelvic cavity, urogenital strategy, and rectum.
The comprehensive scoring criteria were modified from a very similar reporting method on a rat peritoneal carcin omatosis model and set forth in our previous re port. Immunohistochemistry study Tumor tissues obtained from animals of three groups have been subjected to immunohistochemistry to detect the ex pressions of Cat B, Ki 67, CD34, VEGF, E cadherin and D2 40, in accordance to our previously reported procedures.