ABSTRACT
Objective To observe the inhibitory effect on lymph node metastasis of pancreatic cancer and lymphangiogenesis in mice by injection of KAll gene within xenograft tumor.Methods Pancreatic cancer cell line MiaPaCa-2 wag used to construct the nude mice models bearing tumors,then the mice were divided into normal saline group,Ad group and Ad-KAI1 group.Since the successful model construction,normal saline,Ad,Ad-KAI1 was injected every week for 3 times,respectively in the three groups,then the tumor size was documented.50 d after model construction,the tumor and enlarged lymph nodes were collected and subjected to pathological exam,and the expression of LYVE-1 and the MLVD in xenograft tumor was detected by immunohistochemistry.Results Two weeks after MiaPaCa-2 implantation,the model was 100％ successfully constructed.The growth curve of subcutaneous tumor among 3 groups was not statistically significant (P＞0.05) ; the weights of subcutaneous tumor in the 3 groups were (2514.4 ±351.3),(2466.1 ± 295.5),(2294.4±255.4) mg after 50 d,and the difference among the 3 groups was not statistically significant (P ＞0.05).Enlarged lymph nodes metastasis was observed in 8 mice (80％) in normal saline group,and 20 lymph nodes were collected,with 2.0 lymph nodes per mice; and enlarged lymph nodes metastasis was observed in 7 mice (70％) in Ad group,and 15 lymph nodes were collected,with 1.5 lymph nodes per mice; while enlarged lymph nodes metastasis was observed in 4 mice (40％) in Ad-KAI1 group,and 6 lymph nodes were collected,with 0.6 lymph nodes per mice.All the lymph nodes were confirmed to be metastasis of the primary tumor after pathologic exam.The difference of lymph nodes metastasis,number of lymph nodes metastasis per mice among the 3 groups was statistically significant (F =3.14,3.35,P ＜ 0.05).The MLVD of subcutaneous tumor among the 3 groups was (18.70 ± 5.60),(19.40 ± 4.58),(9.80 ±4.10)/400 times magnification,the MLVD of Ad-KAI1 group was significantly lower than those in normal saline group and Ad group (F10.76,11.36,P ＜ 0.05),but the difference between normal saline group and Ad group was not statistically significant.Conclusions KAI1 can inhibit the lymph node metastasis of pancreatic cancer,and the mechanism may be related with decreased lymphangiogenesis and reduced lymphatic vessel density.
ABSTRACT
Objective To investigate the effects of KAll gene transfection on proliferation,migration,invasion and VEGF expression of pancreatic cancer MiaPaCa-2 cells under hypoxic condition,and explore possible mechanism.Methods The pcMV-KAI1 vector which contained the full length of KAI1 cDNA was transfected into pancreatic cancer cells MiaPaCa-2,and KAI1,VEGF C,VEGF A protein expressions were determined by Western blot.The proliferation of pancreatic cancer cells was evaluated by MTT method.Wound-healing assay and cell invasion assay were used to detect the migration and invasion of pancreatic cancer cells.The expression of VEGF C,VEGF A in supernatant of culture was measured by ELISA method.Results The expression of KAI1 protein in MiaPaCa-2 K transfected with KAI1 was significantly higher than that in nontransfected cells [(0.549 ± 0.021) vs 0,P＜0.05].The proliferation under hypoxic condition was not significantly different,but the migration distance was significantly shorter and the number of transmembrane cell was significantly decreased [(14.0 ± 5.8) vs (43.0 ± 14.4),P ＜ 0.05].The expression of VEGF-C in cell was significantly decreased [(0.218 ± 0.043) vs (0.745 ± 0.069),P ＜0.05],but the expression of VEGF-A was not significantly different; the expression of VEGF-C in cell culture supernatants was significantly decreased [(1236 ± 247) vs (2045 ± 221) pg/ml,P ＜ 0.01].Conclusions The migration,invasion ability of pancreatic cancer MiaPaCa-2 cells with KAll transfection under hypoxic condition is decreased,and the possible mechanism of inhibiting lymphatic metastasis is down-regulation of VEGF-C expression.
ABSTRACT
Objective To observe the inhibitory effect on metastasis and growth of pancreatic cancer in mice by injection of KAI1 gene in vivo. Methods Pancreatic cancer cell line MiaPaCa Ⅱ was used to construct the nude mice models bearing tumors, then the mice were divided into normal saline group, Ad group and Ad-KAI1 group. Since the 10th days of model construction, the Ad-KAI1 was injected every 7 d and repeated twice, then the tumor size, the weight of liver, lung and their pathologic changes were evaluated. Results The tumor sizes were not significantly different between the three groups. The weight of lung and liver of Ad-KAI1 group was (0.366±0.041) g and (1. 35±0.21) g, respectively; the weight of lung and liver of Ad group was (0.57±0.065) g and (1.58±1.828) g, respectively; the weight of lung and liver of control group was (0.66±0.13)g and (1.95±0.344)g, respectively. The difference between Ad-KAI1 group and control group was significantly different (t = 5.984, P < 0. 05), and there was no significant difference between Ad group and control group (t=1.089, P > 0.05). The number of pulmonary, liver and lymph node metastasis in Ad-KAI1 group was (1±1), (2±1) and (2±2), respectively; in Ad group was (6±2), (5 ±1), (10±2), respectively; in control group was (7±2), (6±2), (11±3), respectively. The difference between Ad-KAI1 group and control group was significantly different (t = 7.44, 4.34, 8. 16, P < 0.05), while the difference between Ad group and control group was not significantly different (t=0.92, 0.64, 0.42, P >0.05). Conclusions KAI1 gene directly injected into tumors of nude mice may inhibit the growth and metastasis of pancreatic cancer.
ABSTRACT
Objective: To investigate the relationship between the expression of KAI 1 gene and the carcinogenesis and development of thyroid papillary carcinoma (PTC). Methods: The expressions of KAI1 mRNA and protein were detected by reverse transcriptase polymerase chain reaction (RT-PCR) and immunohistochemical method (Envision™) in 59 cases of malignant and benign thyroid tissues, respectively. Their correlation with clinical pathological parameters were analyzed. Results: The level of KAI1 mRNA expression was 3.59 ± 1.57 in PTC, and the expression of KAI1 protein was positive in 24 cases (66.67%). Both the mRNA and the protein expressions of KAI1 were significantly higher than those of follicular adenoma (P <0.01, P <0.05), multinodular goiters (P < 0.01, P < 0.01) and normal thyroid tissues (P < 0.01, P < 0.01). The expressions of KAI1 mRNA and protein had significant correlation with the lymph node metastasis and the tumor size of patients with thyroid papillary carcinoma (P < 0.05). The correlation between KAI1 mRNA and protein are significant positive (r = 0.486, P < 0.01). Conclusion: The abnormal expression of KAI1 mRNA and protein are related with the carcinogenesis and development of thyroid papillary carcinoma. It provides the basis for evaluating the lymph node metastasis and prognosis of thyroid papillary carcinoma.
ABSTRACT
Objective To investigate the effect of recombinant adenovirus containing CD82/KAI1 on the expressions of integrin ?5,integrin ?3,MMP-9,E-cadherin and ?-catenin in uterine stromal cells.Methods Uterine stromal cells were isolated from pregnant mice during implantation window and cultured primarily.The recombinant adenovirus containing mouse CD82/KAI1 gene was constructed.Protein expression of integrin ?5,integrin ?3,MMP-9,E-cadherin and ?-catenin were detected by immunocytochemical methods in the cultured cells with or without CD82/KAI1 adenovirus transfection.Results When the cells which were cultured for 48 h were transfected with the recombinant adenovirus vector at a titer of 6.5?1012 pfu/ml,CD82/KAI1 were obviously upregulated in the primary uterine stromal cells.In the uterine stromal cells from pregnant mice on the 4 d after gestation,integrin ?5,E-cadherin,?-catenin and MMP-9 were detected,while,integrin ?3 were not.But in the cells of nonpregant rats,no expression of these 5 proteins was found.Compared with the cells transfected with blank adenovirus,the recombinant adenovirus encoding CD82/KAI1 upregulated the expressions of ?-catenin and MMP-9(P0.05)and no integrin ?3 was detected in the stromal cells of pregnant mice on day 4 of gestation.Conclusion The recombinant adenovirus containing mouse CD82/KAI1 gene is successfully constructed.CD82/KAI1 might have certain effect on the expressions of integrin ?5,MMP-9,E-cadherin and ?-catenin in mouse uterine stromal cells before embryo implantation.
ABSTRACT
KAI1 gene is a metastasis suppressor gene discovered in 1995, the product of which is a member of the transmembrane-4 superfamily. The expression of KAI1 gene correlate with metastasis of almost all cancer. But some results of some study are different. This article reviewed the study of the genomic structure of KAI1 gene and the relationship between KAI1 gene and neoplastic invasion, metastasis, prognosis.
ABSTRACT
PURPOSE: KAI1/CD82 gene is a recently identified metastasis suppressor gene on human chromosome 11p11.2. Alteration to or reduction of this molecule may allow tumor cells to invade the surrounding tissue and blood vessels. Decreased KAI1 expression seems to be involved in the progression of human prostate, lung and possibly breast cancer, and recently has been demonstrated in several colorectal cell lines. The aim of this study is to determine whether the gene is altered to investigate it in the progression and metastatic process of rectal carcinoma. In addition, its prognostic significance is also evaluated. METHODS: Total 108 tumor samples from primary, metastatic rectal carcinoma were prepared for immunohistochemical study with an anti-KAI1 polyclonal antibody. To analysis the correlation between KAI1 expression and clinicopathological parameter and to evaluate for relation expression and survival. RESULTS: Decrease of KAI1 protein expression was associated with the depth of invasion of tumor (P < 0.0001) and node metastasis (P < 0.05). Liver metastasis showed reduced KAI1 expression when compared with their corresponding primary tumor. Although there was a trend for deteriorating survival from patients with KAI1-positive tumors to those with KAI1-decreased and -negative tumors, it was not significant statistically (P Subject(s)
Humans
, Kangai-1 Protein
, Blood Vessels
, Breast Neoplasms
, Cell Line
, Chromosomes, Human
, Down-Regulation
, Genes, Tumor Suppressor
, Liver
, Lung
, Neoplasm Metastasis
, Prostate
, Rectal Neoplasms
ABSTRACT
Objective To evaluate the effect of recombinant expressing plasmid pCMV-KAI1 on the proliferative ability of PANC Ⅰ and MiaPaCa Ⅱ pancreatic cancer cells, and to observe the suppress metastatic mechanism of KAI1 gene in the malignancy.Methods The plasmid pCMV-KAI1 was transfected into the human pancreatic cancer cell lines PANC Ⅰ and MiaPaCa Ⅱ with liposome. The proliferative ability of these two pancreatic cancer cell lines were analyzed with MTT and colony-forming test, the cycle pattern was assayed by flow cytometry.Results After KAI1 expressing plasmid tranfected into PANC Ⅰ and MiaPaCa Ⅱ, the cell growth rates of the two cell lines were reduced by 40.59% and 65.84% respectively compared with the control cells (P