| Prostate cancer (PCa) is the most common malignant tumor worldwide among older men. In the United States, it has been stood in the first place of new malignant cases for many years and also threatened to men’s health seriously. Due to lack of specific clinical symptoms in early stage, PCa is easily confused with benign prostatic hyperplasia, When patients go to hospital due to symptoms, they usually were diagnosed in the advanced stage, even at terminal of the disease. So, early diagnosis plays a vital role for treatment of PCa. It is well known that serum PSA testing and digital rectal examination are the criterion for prostate cancer screening, but those easily bring about excessive diagnosis and excessive therapy which lead to controversy by many urologist worldwide recently. Although the development of imageology contribute to the diagnosis of prostate cancer, such as MRI spectrum analysis, the golden standard of prostate cancer diagnosis still fix on prostatic biopsy. Given that the risk of trauma, low positive incidence, high infective incidence and potential complication, the biopsy is not satisfied for diagnosis of prostate cancer.As for the treatment of PCa, a variety of effective approaches have achieved great progress on prolonging survival time and improving quality of life, such as radiotherapy, endocrinotherapy, chemotherapeutics. In respect to the mechanism of the occurrence and development of prostate cancer, there are achieved little progresses. Once disease enter the stage of castration-resistant prostate cancer (CRPC), it is failure to offer better choices for patients of PCa in the treatment and prognosis, meanwhile result in death quickly. So, exploring a tumor marker of PCa is essential approach for researching PCa.In the field of cancer research, ion channels keep consistent attention in past decade. A series of ion channels have found participate in oncogenesis. Recent data has shown that transient receptor potential melastatin 7 (TRPM7), a member of transient receptor potential cation channel family (TRP channels), has became a highlight of exploring tumor, evidence has emerged that TRPM7 protein was expressed significantly in human thyroid cancer, breast cancer, gastric cancer, pancreatic cancer, retinoblastoma, lung cancer, etc. So, TRPM7 is considered as significance of some cancers on prognostic guidance and targeted therapy.Meanwhile, what kind of role does TRPM7 play in the occurrence and development of PCa, some explore were carried out worldwide. This present study focus on regulatory role and mechanisms of TRPM7 in PC-3 cells, to evaluate the effect on PC-3 cells treated by TRPM7, and to discuss the signal pathway of TRPM7 in PC-3 cells.The main contents are divided into six sections, as follows:1. TRPM7 was expressed in tissue of prostate cancerIn order to detect the expression of TRPM7 in tissue of prostate cancer, Western blot were used to detect the changes of TRPM7 protein level. Results showed that TRPM7 was increased significantly in tissue of prostate cancer compared with normal prostatic tissue. 2. Inhibiting effect of TRPM7 by Non-specific TRPM7 inhibitor and TRPM7-siRNA in PC-3 cellsNon-specific TRPM7 inhibitor Gd3+ or 2-APB were used to block the expression of TRPM7 in PC-3 cells, we assay the expression of TRPM7 by RT-PCR and Western blot in order to evaluate the effect of expression of TRPM7. Results showed that compared with normal PC-3 cells, Gd3+ or 2-APB decreased TRPM7 mRNA and protein expression in PC-3 cells.Meanwhile, TRPM7-siRNA was transfected to PC-3 cells by Lipofectamine TM2000, the result was also confirmed that expression of TRPM7 was decreased in PC-3 cells.3. Effect of apoptosis by TRPM7 in PC-3 cellsRT-PCR、Western blot and flow cytometry were used to test the apoptosis effect in PC-3 cells by Non-specific TRPM7 inhibitor Gd3+ or 2-APB and TRPM7-siRNA. Results showed that apoptosis of PC-3 cells were increased significantly by treating with Non-specific TRPM7 inhibitor Gd3+ or 2-APB and TRPM7-siRNA. Meanwhile, DR4、 DR5 and cleaved Caspase-3 were also found increased significantly.4. Effect of TRPM7 on PC-3 cells apoptosis induced by TRAILRT-PCR、Western blot and flow cytometry were used to test the effect of TRPM7 in PC-3 cells induced by TRAIL. Results showed that TRPM7 mRNA and protein of actived PC-3 cells were increased significantly compare with the non-actived PC-3 cells.5. Effect of TRPM7 silencing in PC-3 cells apoptosis induced by TRAILRT-PCR、Western blot and flow cytometry were used to assay apoptosis effect on PC-3 cells induced by TRAIL with Non-specific TRPM7 inhibitor Gd3+ or 2-APB and TRPM7-siRNA. Results showed that Non-specific TRPM7 inhibitor Gd3+ or 2-APB and TRPM7-siRNA can promote apoptosis significantly on TRAIL-treated PC-3 cells. Meanwhile, DR4, DR5 and cleaved Caspase-3 were found increased significantly6. Regulatory mechanism of TRPM7 in PC-3 cellsRT-PCR and Western blot were applied to assay the regulation of TRPM7 through AKT pathway in PC-3 cells. Results showed that AKT phosphorylation level was decreased after blocking TRPM7, Meanwhile, downstream protein of AKT 70S6K phosphorylation level was also decreased while blocking TRPM7. The data confirmed that TRPM7 regulate AKT pathway positively in PC-3 cells induced by TRAIL.The above results show that downregulated expression of TRPM7 increased the apoptosis of PC-3 cells. The mechanism may related to AKT signal pathway. The result can offer a novel method for study and treatment of PCa. |
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