Animal Studies of Radio-sensitivity Effects of CK13Gene Mediated by Lipofectin on Nasopharyngeal Carcinoma Cell Strain HNE1
|Keywords||CK13gene NPC radio-sensitivity Animal studies|
Objective:This article discusses the radiosensitive effect of the CK13gene on human nasopharyngeal carcinoma animal model. This topic is on the basis of preliminary research. Animal models of tumors were established in the armpit in Nu-Nu mice with HNE1cell line and HNE1cell line which transferred with target gene. That is NPC animal model group and target gene contained NPC animal model group. In addition, CK13gene expressed plasmid by using polycationic liposome-Mediated was directly injected to NPC animal model group. The controls were injected with empty plasmid and phosphate buffered saline (PBS). Then, all groups were irradiated with a dose of2Gy.4days later tumor injection and radiation were repeated. Observing changes in tumor size before and after treatment and caculating the inhibitory rate. Finally, killing all mice and separating the tumor tissue. Using the method of immunohistochemistry and RT-PCR quantitative to detect the CK13expression. HE staining to compare the tumor tissue necrosis degree of each group. Western blotting testing the CK13protein content. Using CK13as a target, this study investigated the relationship between CK13and NPC radiosensitivity. To privide a new idea for clinical nasopharyngeal carcinoma radiotherapy.Methods:1. The establishment of NPC animal model and target gene contained NPC animal model.2. Injecting plasmid to NPC animal model groups.3. Radiotherapy, and measurement of tumor size before and after radiotherapy.4. Killing all the mice and taking specimens for pathological examination, immunohistochemistry. RT-PCR, and Western Blotting.5. Analyzing the results statistically. Results:In the NPC animal model groups with CK13gene transferred and the groups with CK13plasmid injected, the levels of CK13expression were significantly increased. On the ninth day, when all mice were killed, the inhibition rate of the experimental group (Group A1, A2, B1, B2) were respectively44.6%,47.1%,45.9%, In contrast, the empty vector group (Group C1, C2) inhibition rate was0,0.05%, each experimental group were compared with the empty vector group, the differences were statistically significant (p<0.05). Compared within each experimental group, there was no significant difference. Through immunohistochemical examination, the CK13positive rate of the experimental group was higher than one of the empty vector and contrast group. Besides, the difference was statistically significant. RT-PCR for detection of CK13shows:compared with the empty vector and control groups, the CK13mRNA expression of the experimental group was higher. Western blotting shows: The experimental group appeared significantly CK13protein bands, while the empty vector group and control group, there were no significant bands.Conclusion:CK13gene could increase the sensitivity of NPC to radiation.