| Background and ObjectiveBladder cancer(BC) is the eleventh highest incidence tumor in human all over the world. It ranks as the seventh commonest cancer in male and beyond tenth in female. In the primary diagnosis, about 70%-80% of the bladder cancer are non muscle-invasive bladder cancer, NMIBC, the remaining 20%-30% are muscle-invasive bladder cancer, MIBC. From pathology examination, though both the two kinds of bladder cancers originate from bladder urotherial cells, they obviously present different clinical characteristics. Compared with other human malignant tumors, the overall survival is comparatively good in patients with non muscle-invasive bladder cancer. However, after the primary tumor is resected, 30%-50% of the patients have the possibility of a recurrence and 10%-20% of the patients with NMIBC progress to MIBC. Though only 20% of patients with bladder cancer were diagnosed as MIBC at initial diagnosis, most of those cancer patients progress to death. Besides, the occult distant metastasis have been found among nearly 50% patients with MIBC at initial diagnosis. Therefore, the high risk factors of NMIBC recurrence and progress,and the metastasis and prognosis of patients with MIBC are matters of concern and remained to be solved by us, clinical scientist and experimenter. So early diagnosis and treatment of bladder cancer has become extremely important.Heme-responsive gene 1,HRG-1, found in our university, is a new gene which can induce tumor cell apoptosis. It is the sole member of solute carriers,SLC48, related to endosome. This gene can carry heme from endosome to cytoplasm. The interaction between SLC48A1/HRG-1 and V-ATP enzyme acidfication endosome can increase the enzyme activity of V-ATP as well as the recycling of membrane transporter and receptor. The previous studies in our university suggested that HRG-1 show high expression in the prostatic neoplasms tissues, and low expression or no expression in benign prostatic hyperplasia, thus it provides a theoretical basis for us. There is an interaction between HRG-1 and V-ATP enzyme reported in literature, V-ATP enzyme have participated in the tumor occurrence and invasion, thus inform us the recurrence of bladder neoplasm.The purpose of the present research is to explore the expressions of HRG-1 in bladder cancer and normal bladder tissues, to transfect the target cells by using si RNA interference technique in order to regulate the expression of HRG-1 protein in bladder cancer cells, to examine the effects of observation period on the proliferation of human bladder cancer cell so as to provide theoretical basis for clinical work and cancer prognosis.Methods1.To observe the expression of HRG-1 in normal bladder tissues and in different grade bladder cancer tissues, and find out whether HRG-1 gene can provide the possible guidance for the diagnosis and treatment of patients with bladder cancer.2. Three groups of bladder cancer cells were selected by western blot method and real-time PCR, the group of bladder cancer cells in which the expression of HRG-1 was the highest was available for the research.3.The HRG-1 si RNA was transfected into bladder cancer T24 cell lines via RNAi, and measure the expression changes of HRG-1 si RNA in bladder cancer T24 cell lines by real-time PCR and western blot method.4. Interfere the expression of HRG-1, and detect the effect on proliferation and apoptosis for bladder cancer T24 cell lines by MTT assay and flow cytometry.Results1.The expression of HRG-1 was obviously stronger in bladder cancer tissues than in normal bladder tissues with the positive rate 71.8% and 25% respectively. And the expression of HRG-1 in bladder tissues was not related to the sex and age, but was positively correlated with the pathological stage of tumor.2. The expression of T24 cell, 5637 cell, and BIU-87 cell was analyzed by western blot method and real-time PCR. The results showed that the expression of HRG-1was the highest in bladder cancer T24 cell.3.The HRG-1 si RNA gene was successfully transfected into bladder cancer T24 cell lines via RNAi. The results showed that compared with control group, the expression of HRG-1 si RNA in bladder cancer T24 cell lines was significantly down.4.MTT assay and flow cytometry detection showed that compared with the control group and blank group, the proliferation and apoptosis of T24 cell lines in HRG-1 si RNA experimental group was decreased significantly.Conclusions1.There was higher expression of HRG-1 in bladder cancer tissues and less or no expression of HRG-1 in normal bladder tissues, which suggested that the expression of HRG-1 is closely related to the grade and tumor progression. The results provide the basis for the diagnosis and treatment of bladder cancer.2.HRG-1 show differential expressions in different bladder cancer cells. The downregulated expression of HRG-1 in bladder cancer T24 cell can inhibit the proliferation and metastasis of T24 cell, indicating that HRG-1 may involve in the development and progression of cancer. |
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