The Effect Of TSLC1 On Proliferation, Invasion And Metastasis Of Human Prostatic Carcinoma Cell Line T₃B

Objective 1. To establish the prostatic carcinoma cell subline which expressingTSLC1 protein and to identify its biological characteristics. 2. To elucidate the effect ofTSLC1 on proliferation and invasion of human prostatic carcinoma cell line T₃B. 3. Tostudy the effect of TSLC1 on tumorigenicity and metastasis of human prostatic carcinomacell line T₃B.Methods 1. The recombinant plasmid pCI-TSLC1 was stably transfected into T₃Bcells with Lipofectamine 2000. The positive clones were developed by selection by G418.Biological characteristics of T₃B cells which were stably transfected with pCI-TSLC1 werestudied. 2. The T₃B cells stably transfected with exogenous gene TSLC1 (experimentalgroup) and those treated with pCI-neo vector (control group) and without any treatment(blank group) were compared. Cell proliferation was analyzed with MTT assay. FACSortflow cytometry analysis was performed to assess the cell cycle distribution and apoptosis.Transwell was performed to assess the ability of invasion. The speed of cell migration wasdetected by cell scratch method. 3. The experimental group, control group and blank groupcells were injected respectively into nude mices subcutaneously with cell concentration of4.0×10⁶/200μL. The growth of tumor was examined in three groups. The three groups cellconcentration of 4.0×10⁶/200μL was injected into nude mice through vena caudalis toobserve metastasis. The three groups cell concentration of 2.0×10⁶/10μL were injectedin-situ to establish osseous metastasis model. We examined the rate of osseous metastasis inthree groups.Results 1. The stable cell subline highly expressing TSLC1 protein was obtained. The ectogenous gene TSLC1 has integrated into the genomes of T₃B cells which wereidentified from both nucleic acid and protein levels. The genetic stability and purity of thecell population were confirmed. 2. The growth of TSLC1-transfected cells was significantlysuppressed in vitro compared with those of the control and blank groups, displaying that theamount of G₀/G₁ cells increased and the amount of S phase cells decreased significantly,which suggest a G₀/G₁ cell cycle arresting. The number of cells in early, late and total phaseapoptosis were significantly higher than those of the control and blank groups (P＜0.01).The ability of invasion and migration were significantly suppressed compared with those ofthe control and blank groups (P＜0.05). 3. The subcutaneous tumor of the experimentalgroup occurred later and smaller than the control and blank groups(P＜0.01). Lungmetastases of nude mice occurred later and less than the control and blank groups. The rateof osseous metastasis was 20% in experimental group, which decreased significantly(P＜0.05) compared with the control group (100%) and the blank group (100%).Conclusions 1. The stable prostatic carcinoma cell subline highly expressingTSLC1 has successfully established, which provided a basis for further exploring the rolesof TSLC1 in prostatic carcinoma. 2. TSLC1 strongly inhibits the proliferation and invasionof T₃B cells in vitro and induces apoptosis of cells. 3. TSLC1 strongly inhibits thetumorigenicity and metastasis of T₃B cells...