Objective To discuss the feasibility on building lewis lung carcinoma mouse models through different methods and improve the methods. Methods The method of culture LLC cells in vitro, trypsin digestion method, Ⅳ collagenase method and homogenate method were compared to make the different dose of cell suspension injected into C57BL/6 mice. The feasibility of the improved method was determined through observing the cell count, the tumor formation ratio, the tumor formation time, tumor volume, weight and life habit. Results The method of culture LLC cells in vitro could get needed cells and its tumor formation ratio was 100 %. Trypsin digestion method and homogenate method could get less cells and its tumor formation ratio was about 80 %～90 % and 60 %～75 %. Whereas 1V collagenase method could get most cell count and its tumor formation ratio was 100 %. Conclusion IV collagenase method is a preferred method which is simple,high efficiency and make a strong base on the cancer experimental study.

Objective To explore the specific cellular and humoral immunity induced by dendritic cells (DC) vaccine loading allogenic microvascular endothelial cell bEnd. 3 antigen against U14 cervical cancer cell of mice. Methods Mouse brain microvascular endothelial cell bEnd. 3 was cultured and identified for preparation endothelial cell bEnd. 3 antigen. The level of mRNA expression of vascular endothelial growth factor receptor 2 (VEGF-R2) and integrin αV was detected by reverse transcription (RT)-PCR. The BALB/c mice were immuned with DC loading bEnd. 3 antigen 4 times in 4 weeks (bEnd. 3-DC group), while the mice only were immuned with DC or injected with phosphate buffer saline (PBS group) as control group. One week after last vaccination, U14 cervical cancer cells were injected subcutaneously into the mice. The tumor size, cytotoxic T lymphocyte (CTL) response of spleen lymphocytes in vitro, the percentage of CD3+ CD+8 surface markers of spleen lymphocytes, and the titer of serum antibody were detected. The specific immunity was examined by immunocytochemistry and western blot. Results The expression of VEGF-R2 and integrin αV gene in bEnd. 3 cells were expressed highly.After the vaccine was injected, the tumors of mice in PBS group grew faster than those in other groups, while the tumors in bEnd. 3-DC group grew slowly and disappeared after 2 weeks. The volume of tumors in DC group grew slower than those in PBS group [(0.11± 0.13) cm3 versus (3.38 ±0.34) cm3]. The CTL response of spleen iymphocytes in vitro showed that bEnd. 3-DC cells could kill bEnd. 3 cells, the special lysis rate was more than 60% . The percentage of CD+3 CD+8 spleen lymphocytes in bEnd. 3-DC group[(38.6 ± 0.7) %] was higher than those in other groups (P ＜ 0.05). The titer of serum antibody of Immunocytochemistry analysis indicated there were specific antigen-antibody reaction to bEnd. 3 cell in bEnd. 3-DC group. Western blot analysis revealed that there were specific bands at 220 000 (VEGF-R2).Conclusions bEnd. 3-DC vaccine can inhibit the tumor growth of U14 cervical cancer cell of mice, which indicates that the special cellular and humorai immunity are induced by bEnd. 3-DC antigen which maybe have some antigens in bEnd. 3 cells that reacts with endothelial cell proliferation-related antigens.

Objective To study the expression of kinesin family member C1 ( KIFC1 ) in hepatocellular carcinoma (HCC) and analyze its correlation with the clinicopathological features and prognosis of patients. Methods The expression levels of KIFC1 protein in the HCC tissues from 82 patients were determined by immunohistochemical staining. The correlation between KIFC1 protein and clinicopathological characteristics (including age, gender, tumor nodules, tumor grade, tumor volume, lymph node metastasis, and alpha-fetoprotein expression) was analyzed. The Kaplan-Meier analysis was used to analyze the effect of KIFC1 expression level on overall survival and progression-free survival in patients with HCC. The expression level of KIFC1 mRNA in liver cancer tissue was analyzed by GPEIA database. The correlation between KIFC1 expression and prognosis was analyzed by KM-plotter. Results KIFC1 protein is significantly overexpressed in liver cancer tissues, and its expression level is significantly correlated with tumor nodule number (P=0.023) and tumor size (P=0.011). Patients with high expression of KIFC1 had poor overall disease and disease-free survival (all P<0.05). KIFC1 mRNA is significantly overexpressed in liver cancer tissues and correlated with disease-free survival and overall survival. Conclusions The expression of KIFC1 protein is highly expressed in liver cancer tissues, and its expression level is related to the clinicopathological characteristics of liver cancer. Bioinformatics analysis results show that KIFC1 is related to the poor prognosis of patients, suggesting that KIFC1 is expected to be a potential predictor and therapeutic target for liver cancer prognosis.