The effects of over-expression of testis-specific expressed gene 1 (TSEG-1) on the viability and apoptosis of cultured spermatogonial GC-1spg cells were investigated, and the immortal spermatogonial cell line GC-1spg (CRL-2053™) was obtained as the cell model in order to explore the function of TSEG-1. We transfected the eukaryotic vector of TSEG-1, named as pEGFP-TSEG-1 into cultured spermatogonial GC-1spg cells. Over-expression of TSEG-1 inhibited the proliferation of GC-1spg cells, and arrested cell cycle slightly at G0/G1 phase. Transfection of TSEG-1 attenuated the transcript levels of Ki-67, PCNA and cyclin D1. In addition, over-expression of TSEG-1 induced early and late apoptosis, and reduced the mitochondrial membrane potential of GC-1spg cells. Moreover, transfection of TSEG-1 significantly enhanced the ratio of Bax/Bcl-2 and transcript levels of caspase 9, and decreased the expression of Fas and caspase 8 in GC-1spg cells. These results indicated over-expression of TSEG-1 suppresses the proliferation and induces the apoptosis of GC-1spg cells, which establishes a basis for further study on the function of TSEG-1.
Animals ; Caspase 8 ; biosynthesis ; genetics ; Cell Line ; Cyclin D1 ; biosynthesis ; genetics ; G1 Phase ; physiology ; Histones ; genetics ; metabolism ; Ki-67 Antigen ; biosynthesis ; genetics ; Male ; Mice ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Resting Phase, Cell Cycle ; physiology ; Spermatogonia ; cytology ; metabolism ; bcl-2-Associated X Protein ; biosynthesis ; genetics

OBJECTIVETo study the effect of pGCsi-H1-APRIL on the growth of human colorectal cancer cells in transplated tumor in nude mice and to improve the effect of APRIL on proliferation and apoptosis of colorectal cancer (CRC).

METHODSHuman CRC model was established in nude mice, and the nude mice were treated with APRIL siRNA twice per week for 2 weeks. APRIL mRNA expression was surveyed by PCR and APRIL protein expression was detected by immunohistochemistry. The expression of PCNA protein was detected by ELISA. The expression of bcl-2 and bcl-xl was assessed by immunohistochemical staining, and TUNEL staining was used to detect apoptosis.

RESULTSThe expression of APRIL mRNA in the APRIL siRNA group was (0.13 ± 0.05) × 10(-3), significantly lower than that in the vector group (0.95 ± 0.04) × 10(-3) and the PBS group (0.96 ± 0.05) × 10(-3). The expression of APRIL protein in the APRIL siRNA group was (87.5 ± 5.0)% lower than that in the vector and PBS groups (P < 0.05). APRIL siRNA significantly suppressed the growth of SW480 tumor: the IR (inhibitory rate) of APRIL siRNA group was (60.7 ± 1.5)% (P < 0.05). The expression of PCNA in APRIL siRNA group was (176.8 ± 18.1) ng/ml, was (56.5 ± 2.0)% lower than that of PBS group (328.4 ± 22.8) ng/ml. Furthermore, the expressions of anti-apoptosis proteins bcl-2 and bcl-xl of APRIL siRNA group were (82.6 ± 4.5)% and (79.2 ± 3.5)% lower than those of the PBS group. The apoptotic rate of the APRIL siRNA group was 40.1% ± 2.5%, significantly higher than that in the vector group (2.5 ± 0.1)% and PBS group (2.5 ± 0.2)% (P < 0.05).

CONCLUSIONAPRIL siRNA may significantly suppress the growth and promote apoptosis in transplanted tumor of human colorectal cancer in nude mice. APRIL may become a candidate gene of gene therapy of human colorectal cancer.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Colorectal Neoplasms ; genetics ; metabolism ; pathology ; Female ; Humans ; Ligands ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proliferating Cell Nuclear Antigen ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Random Allocation ; Tumor Necrosis Factor Ligand Superfamily Member 13 ; biosynthesis ; genetics ; bcl-X Protein ; metabolism

Even though there is no direct evidence to prove the cellular and molecular changes induced by radiofrequency (RF) radiation itself, we cannot completely exclude the possibility of any biological effect of mobile phone frequency radiation. We established a carousel-type exposure chamber for 849 MHz or 1763 MHz of mobile phone RF radiation to expose RF to the heads of C57BL mice. In this chamber, animals were irradiated intermittently at 7.8 W/kg for a maximum of 12 months. During this period, the body weights of 3 groups-sham, 849 MHz RF, and 1763 MHz RF-did not show any differences between groups. The brain tissues were obtained from 3 groups at 6 months and 12 months to examine the differences in histology and cell proliferation between control and RF exposure groups, but we could not find any change upon RF radiation. Likewise, we could not find changes in the expression and distribution of NeuN and GFAP in hippocampus and cerebellum, or in cell death by TUNEL assay in RF exposure groups. From these data, we conclude that the chronic exposure to 849 MHz and 1763 MHz RF radiation at a 7.8 W/kg specific absorption rate (SAR) could not induce cellular alterations such as proliferation, death, and reactive gliosis.
Animals ; Apoptosis/*radiation effects ; Body Weight/radiation effects ; Brain/pathology/*radiation effects ; Cell Proliferation/*radiation effects ; *Cellular Phone ; Dose-Response Relationship, Radiation ; Gliosis/etiology/pathology ; In Situ Nick-End Labeling ; Mice ; Mice, Inbred C57BL ; Nerve Tissue Proteins/biosynthesis/genetics ; Proliferating Cell Nuclear Antigen/biosynthesis/genetics ; Radio Waves/*adverse effects

OBJECTIVE: To investigate the expression of Skp2 and its relation with P27 expression, clinic pathologic features, and prognostic indicator in osteosarcoma. METHODS: We collected osteosarcoma specimen from 52 patients (29 males and 23 females), who were all treated by radical resection of tumor. The expression of Skp2 and P27 was determined by SP immunohistochemistry. Forty-four patients were followed up for 4 to approximately 84(mean = 31.2)months, while the other 8 patients were lost. Twenty of them survived over 5 years and 24 died. RESULTS: In osteosarcoma, Skp2 highly expressed (mean value was 1.74). Expression intensity of Skp2 at the stage III was obviously higher than that of the stage II(IIa and IIb) (P < 0.05). Skp2 expression was correlated with the relapse, metastasis, and 5-year survival in osteosarcoma (P < 0.05), but not with different pathologic types, sex, or age(P > 0.05). The expressions of skp2 and P27 were negative correlation in osteosarcoma (r = -0.907, P < 0.05). CONCLUSION Skp2 plays an important role in the occurrence and development of osteocarcoma by causing the degradation of P27, and can be an important prognostic indicator in osteosarcoma.
Adolescent ; Adult ; Biomarkers, Tumor ; Bone Neoplasms ; metabolism ; pathology ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Osteosarcoma ; metabolism ; pathology ; Prognosis ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; S-Phase Kinase-Associated Proteins ; biosynthesis ; genetics ; Tumor Cells, Cultured

To investigate the role of progesterone receptor (PR) expression in malignant melanoma (MM), PR and proliferative cell nuclear antigen (PCNA) expression were immunohistochemistrically evaluated in a series of 35 specimens of MM, and the correlation between the immunohistochemistrical findings and clinicopathological data was also analyzed. PR expression was detected in 25.7% (9/35) of the patients with MM. No PR expression was observed in nevi. PR expression was inversely correlated with PCNA expression (r=-0.353, P=0.026). PR expression was slightly increased in females, subjects aged under 55 y, those with ulceration, non-acral subtype and diagnosis delay longer than 1 y, but the difference was not statistically significant. Selective expression of progesterone receptor in malignant melanoma might be correlated with inhibited tumor growth.
Gene Expression Regulation, Neoplastic ; Immunohistochemistry ; Melanoma/*metabolism ; Models, Biological ; Prognosis ; Proliferating Cell Nuclear Antigen/*metabolism ; Receptors, Progesterone/*biosynthesis ; Receptors, Progesterone/genetics ; Skin/metabolism ; Skin Neoplasms/metabolism

The study was aimed to explore the possible roles of survivin and P63 protein in the development and progression of B cell non-Hodgkin's lymphoma (B-NHL) and their relation with cell apoptosis and proliferation. TdT-mediated dUTP nick end labeling (TUNEL) and immunohistochemistry were used to detect the survivin and P63 protein expression, cell apoptosis and proliferating cell nuclear antigen (PCNA) level in 43 cases of B-NHL and 10 cases of reactive hyperplasia lymphoid (RHL) tissues. The results indicated that the positive rates of survivin and P63 protein expression were 69.8% (30/43) and 82.7% (30/43) respectively. The expression of survivin and P63 protein was 10% (1/10) and 40% (4/10) in RHL tissues of 10 cases. The expression of survivin in aggression B-NHL was higher than that in indolent B-NHL (83.3% vs 46.2%, P < 0.01). The expression of P63 proteins in aggressive B-NHL was higher than that in indolent B-NHL (86.7% vs 76.9%, P > 0.05). Apoptotic index (AI) and proliferation index (PI) correlated positively with expression of survivin (r = 0.429, P < 0.01; r = 0.348, P < 0.01), and so do with expression of P63 proteins (r = 0.451, P < 0.01; r = 0.369, P < 0.05). In addition, AI and PI were positively related (r = 0.598, P < 0.001). It is concluded that survivin may participate in the regulation mechanism of B-NHL cell apoptosis and proliferation, P63 as an oncogene enhances proliferation and takes part in the development of B-NHL. There may be a close relationship between survivin and P63 protein in the regulation of lymphocyte proliferative kinetics.
Adolescent ; Adult ; Aged ; Apoptosis ; genetics ; Cell Proliferation ; Child ; Child, Preschool ; DNA-Binding Proteins ; biosynthesis ; genetics ; Female ; Humans ; Inhibitor of Apoptosis Proteins ; Lymphoma, B-Cell ; metabolism ; Male ; Microtubule-Associated Proteins ; biosynthesis ; genetics ; Middle Aged ; Neoplasm Proteins ; biosynthesis ; genetics ; Proliferating Cell Nuclear Antigen ; metabolism ; Trans-Activators ; biosynthesis ; genetics ; Transcription Factors ; Tumor Suppressor Proteins ; biosynthesis ; genetics

OBJECTIVETo determine and compare the difference of cell apoptosis and proliferation in the transition and peripheral zones in the human prostate.

METHODSSeventeen normal prostate glands from organ donors were sampled from normal men according to McNeal/s zonal anatomy, and 20 hyperplastic transition zones obtained from prostatectomy specimens of BPH patients. Cell proliferation and Bcl-2 expression were assessed by immunostaining using PCNA and anti-Bcl-2 antibodies, while apoptotic bodies were specifically stained using TUNEL. Bcl-2 mRNA expression was detected by RT-PCR.

RESULTSIn the normal epithelium, the rates of cell proliferation and apoptosis were markedly decreased in the transition zone as compared with the peripheral zone. The proliferation index was significantly increased in the hyperplastic transition zone in BPH, while the apoptosis index significantly decreased in comparison with the normal prostate. Bcl-2 was significantly greater in the normal transition epithelium than in the peripheral zone, and over-expressed in the hyperplastic transition zone. There was a significant negative correlation between the Bcl-2 expression and the apoptosis of the epithelial cells in the hyperplastic transition zone (r(s) = -0.867, P < 0.01).

CONCLUSIONThe hyperplastic transition zone may result from both an increase of cell proliferation and a failure of cell apoptosis. Increased expression of Bcl-2 may participate in the BPH process by blocking cell apoptosis.

Adult ; Apoptosis ; Case-Control Studies ; Cell Proliferation ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Male ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; Prostate ; cytology ; metabolism ; Prostatic Hyperplasia ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; RNA, Messenger ; genetics

OBJECTIVETo investigate the expressions of human telomerase reverse transcriptase (h-TERT), c-myc, and proliferating cell nuclear antigen (PCNA) in chronic viral hepatitis (CVH), liver cirrhosis and primary hepatocellular carcinoma (HCC) and understand their possible role in liver carcinogenesis.

METHODSTotally 157 liver disease specimens were collected, including 56 CVH, 52 liver cirrhosis and 49 primary HCC specimens. In situ hybridization was performed on these specimens to examine the expressions of h-TRET and c-myc mRNA, and immunohistochemistry carried out for PCNA detection, with the cell apoptosis detected with in situ ending labeling.

RESULTSIn the CVH, liver cirrhosis and primary HCC specimens, h-TERT expression was detected at the frequencies of 11/56 (19.6%), 43/52 (82.7%) and 44/47 (93.6%), c-myc expression at 7/56 (12.5%), 21/52 (40.4%) and 26/47 (55.3%), with apoptotic index of (27.3-/+4.7)%, (16.5-/+2.6)% and (8.7-/+1.3)% and PCNA expression rate of (17.1-/+2.9)%, (49.3-/+7.8)% and (62.5-/+9.1)%, respectively. Correlations among h-TERT, c-myc, and PCNA expressions and the apoptotic index were not found in the examined tissues (P>0.05).

CONCLUSIONLiver carcinogenesis may involve increased h-TERT, c-myc, and PCNA expressions and suppressed cell apoptosis.

Adult ; Apoptosis ; Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Cell Transformation, Neoplastic ; Female ; Hepatitis B, Chronic ; genetics ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Liver Cirrhosis ; genetics ; metabolism ; pathology ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Male ; Middle Aged ; Proliferating Cell Nuclear Antigen ; biosynthesis ; Proto-Oncogene Proteins c-myc ; genetics ; RNA, Messenger ; genetics ; metabolism ; Telomerase ; genetics

OBJECTIVETo dynamically observe the effect of Shenshuai Mixture (SM) on repair of kidney in acute renal failure (ARF) rats.

METHODSMale SD rats were divided into 4 groups randomly, the normal group, the SM group, the verapamil group and the model control group. Except those in the normal group were treated with normal saline without modeling, all remaining rats, after being made into ARF model by intra-muscular injection of glycerin, were treated with SM, verapamil and normal saline respectively via gastrogavage. Renal function, renal pathology, mRNA expression of epidermal growth factor (EGF) and protein expression of proliferating cell nuclear antigen (PCNA) were detected once every day from the 1st day to the 5th day. Results (1) BUN and Scr levels increased markedly 24 hrs after modeling, but the Scr level in the two treated groups was significantly lower than that in the model group (P < 0.05). Compared with that in the model group and the verapamil group, renal function was better in the SM group on the 3rd day (P < 0.01), and approach to normal level on the 5th day. (2) Renal pathological changes alleviated in every phase of ARF in the SM group than that in the model group, especially part of tubule regeneration could be seen on the 3rd day (metaphase), and renal structure was rehabilitated on the 5th day (convalescence), prior to those in the model group. (3) At the 3rd day expression of EGF mRNA and PCNA in tubule epithelial cell (TEC) increased remarkably in the SM group, higher than those in the model and verapamil group (P < 0.05).

CONCLUSIONSM could promote renal tissue regeneration and rehabilitation, and shorten the course of ARF through up-regulating mRNA expression of EGF in TEC.

Acute Kidney Injury ; drug therapy ; physiopathology ; Animals ; Drugs, Chinese Herbal ; therapeutic use ; Epidermal Growth Factor ; biosynthesis ; genetics ; Kidney ; physiopathology ; Male ; Phytotherapy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo investigate the effects and mechanisms of Yishen Huoxue decoction (YHD) on chronic renal failure (CRF) rats induced by 5/6 nephrectomy.

METHODSThe glomerulosclerosis model was established by 5/6 nephrectomy in rats. Experimental animals were allocated into the normal group, the model group, the YHD group and the benazepril group. Urine protein of 24 h (UP) at the 6th and 12th weekend after operation, blood urea nitrogen (BUN) and creatinine (SCr), albumin (Alb) and haemoglobin (HB) at the 12th weekend were measured, renal pathology changes were examined with light microscope, the expressions of proliferating cell nuclear antigen (PCNA) and fibronectin (Fn) were examined by immunohistochemistry and mRNA expressions of connective tissue growth factor (CTGF) and plasminogen activator inhibitor-1 (PAI-1) by RT-PCR at the 12th weekend.

RESULTSCompared with those in the normal group, the levels of UP, BUN and SCr, the area of glomerular mesangial matrix, the FN deposition, PCNA expression in glomeruli and tubular interstitium and mRNA expressions of CTGF and PAI-1 were all significantly higher in the model group (P < 0.05). All the above-mentioned indexes were lower in the YHD group than those in the model group (P < 0.05). PCNA positively expressed cells in glomeruli of the normal, model group, YHD group and benazepril group was 7.00 +/- 2.24,34.78 +/- 6.96,15.75 +/- 2.61 and 15.50 +/- 2.57 respectively, positively correlated to the expression of CTGF, PAI-1, FN and SCr level.

CONCLUSIONYHD could delay the progression of CRF in 5/6 nephrectomized rats, and the mechanisms were mainly related to the inhibition on renal cell proliferation and it induced over-expression of cytokines, and accumulation of extracellular matrix.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Extracellular Matrix ; drug effects ; metabolism ; Fibronectins ; biosynthesis ; genetics ; Glomerulosclerosis, Focal Segmental ; drug therapy ; etiology ; metabolism ; Kidney Failure, Chronic ; drug therapy ; etiology ; metabolism ; Male ; Nephrectomy ; Phytotherapy ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction