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

OBJECTIVE: To investigate the significance of expression of COX-2, p21, Ki67 and HPV in nasal inverted papilloma. METHOD: Detecting COX-2, p21, Ki-67 in 30 cases of nasal inverted papilloma (NIP), 20 cases of nasal polyps (NP) and 10 cases of normal nasal mucosa (NM) by two step immunohistochemical method, and HPV virus by flow-through hybridization method. RESULT: The positive expression rate of COX-2 and Ki-67 in NIP, NP and NM group was decreased in turn, COX-2 had significant difference in the groups(χ2 = 30.00, P< 0. 05); the positive expression rate of Ki-67 had significant differences between NIP and NM group (χ2 = 8. 533, P<0. 05). The expression of COX-2 in NIP tissues was positively correlate with that of Ki-67 by using Spearman rank correlation analysis (r=0.78, P<0.05). Expression of p21 were not observed in NIP group. The positive rate of HPV was 26. 67% in 30 cases of NIP, all of HPV16 type. CONCLUSION COX-2, Ki-67 and HPV infection have certain correlation with the occurrence of NIP. The occurrence of NIP has relationship with inflammatory reaction mediated by COX-2. Ki-67 can well reflect the proliferation activity of tumor cells, and can be used to measure the proliferation rate of nasal inverted papilloma. The COX-2 and Ki-67 have a synergistic role in the pathogenesis of NIP. p21 has no significant relationship with the incidence of NIP. HPV infection is related to the pathogenesis of NIP, but not as a;major factor in the pathogenesis of NIP.
Case-Control Studies ; Cyclin-Dependent Kinase Inhibitor p21 ; biosynthesis ; Cyclooxygenase 2 ; biosynthesis ; Humans ; Ki-67 Antigen ; biosynthesis ; Nasal Mucosa ; Nasal Polyps ; Nose Neoplasms ; genetics ; virology ; Papilloma, Inverted ; genetics ; virology ; Papillomavirus Infections

OBJECTIVE: To investigate the expression and clinical significance of Ki67 and calcitonin in medullary thyroid carcinoma(MTC). METHOD: The expression level of Ki67 and calcitonin was studied in 44 cases of medullary thyroid carcinoma tissue and 20 cases of adjacent nontumor tissue by SP immunohistochemistry. RESULT: The positive expression of Ki67 and calcitonin in medullary thyroid carcinoma tissue were 86.36% (38/44) and 100.00% (44/44) respectively. There was a significant difference between carcinoma and normal thyroid tissue (P<0.01). The overexpression of Ki67 and calcitonin in medullary thyroid carcinoma had no relationship with gender and age of patients,but had relationship with size of tumor,clinical staging and lymph node metastasis (P<0.05). Meanwhile, Ki67 and calcitonon had no significant correlation with each other. CONCLUSION The overexpression of Ki67 and calcitonin may play important role in occurrence, development and metastasis of medullary thyroid carcinoma. It may be used as an important judgement for the biological behavior of medullary thyroid carcinoma.
Calcitonin ; biosynthesis ; Carcinoma, Neuroendocrine ; Humans ; Ki-67 Antigen ; biosynthesis ; Lymphatic Metastasis ; Thyroid Neoplasms ; metabolism

OBJECTIVETo study the inhibitory effect of knocking down microRNA(miR)-221 and miR-222 on human glioma cell growth and its possible mechanism.

METHODSmiRNA-221/222 antisense oligonucleotides (antisense miR221/222) were transfected into human glioma U251 cells by lipofectamine. Northern blot analysis was conducted to detect the mRNA expression of miR-221/222 in the control and transfected cell groups. The proliferation activity of cells was determined by MTT assay. Cell invasion ability was examined by transwell assay, and cell cycle kinetics and apoptosis were detected with flow cytometry. The expression of relevant proteins was analyzed by Western blotting. The therapeutic efficacy of antisense miR221/222 on the growth of xenograft tumors in nude mice were also observed.

RESULTSIn the antisense miR-221/222-transfected cells, the expression of miR-221/222 was significantly reduced; the cell invasion ability was suppressed, cell cycle was blocked at G(0)/G(1) phase, and apoptotic cells were increased. The growth of xenograft tumors treated with antisense miR-221/222 was also inhibited. In antisense miR-221/222 treated tumor cells, the expression of bcl-2 was down-regulated while connexin43, p27, PUMA, caspase-3, PTEN, TIMP3 and Bax up-regulated, and p53 expression not changed.

CONCLUSIONThere is a significant inhibitory effect of antisense miR-221/222 on the growth of human glioma U251 cells. miR-221/222 may be considered as a candidate target for gene therapy of human gliomas.

Animals ; Apoptosis ; Base Sequence ; Caspase 3 ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Gene Knockdown Techniques ; Genetic Therapy ; Glioma ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; metabolism ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; MicroRNAs ; biosynthesis ; genetics ; Molecular Sequence Data ; Neoplasm Transplantation ; Oligonucleotides, Antisense ; pharmacology ; PTEN Phosphohydrolase ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; RNA, Messenger ; metabolism ; Tissue Inhibitor of Metalloproteinase-3 ; metabolism ; Transfection

OBJECTIVE: To detect the expression of Ki-67, Bcl-2, Bax and caspase-3 in simple benign prostatic hyperplasia (BPH) and BPH combined with prostatitis,and to evaluate the effect of inflammation on the development and progression of BPH. METHODS: All specimens were obtained from patients undergoing surgical resection of the prostate. The paraffin section of the specimens was stained with hemotoxyline and eosin, and observed under light microscope to examine the inflammation hispathological changes. Sixteen patients with simple BPH (Group A) and 42 patients with BPH combined with prostatitis (Group B) were included. Immunohistochemical analysis and Western blot were used to examine the expression of Ki-67, Bcl-2, Bax and caspase-3. RESULTS: The expression of Ki-67 and Bcl-2 was significantly higher in Group B than that in Group A (P<0.05), and caspase-3 expression was significantly lower (P<0.05). There was no difference in Bax expression between the 2 groups (P>0.05). CONCLUSION Prostatitis can up-regulate Ki-67, Bcl-2 expression, and down-regulate the expression of caspase-3 in BPH. Prostatitis appeared to play an important role in the development of BPH by affecting the proliferation and apoptosis of the prostatic cells.
Aged ; Caspase 3 ; metabolism ; Humans ; Ki-67 Antigen ; biosynthesis ; Male ; Middle Aged ; Prostatic Hyperplasia ; complications ; metabolism ; Prostatitis ; complications ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; Up-Regulation ; bcl-2-Associated X Protein ; biosynthesis

AIMTo examine the impact and prognostic significance of alpha-tocopherol associated protein (TAP) expression in a series of prostate cancer patients.

METHODSTissues from 87 patients underwent radical prostatectomy were examined for TAP expression by immunohistochemistry. The relationships of the staining results, the clinic pathological characteristics and the recurrence times were analyzed.

RESULTSCompared with the adjacent areas of normal and benign glands, immunoreactivity of TAP was reduced in areas of prostate cancer. A lower TAP-positive cell number per mm(2) of the largest cancer area (defined as TAP-PN) was associated with higher clinical stage (r = -0.248, P = 0.0322). Inverse associations were found among the TAP-PN and positive lymph nodes (r = -0.231, P = 0.0325), preoperative prostate-specific antigen (PSA) levels (r = -0.423, P = 0.0043), tumor size (r= -0.315, P= 0.0210) and elevated tumor cell proliferation, which was indicated by the staining of Ki-67 (r = -0.308, P = 0.0026). TAP-PN was a significant predictor of recurrence univariately (P = 0.0006), as well as multivariately, adjusted for known markers including preoperative PSA, clinical stage, Gleason score, surgical margin, extra-prostatic extension, seminal vesicle invasion and lymph node metastasis (P = 0.0012).

CONCLUSIONReduced expression of TAP was associated with the cell proliferation status of prostate cancer, adverse pathological parameters and the increased risk of recurrence.

Aged ; Carrier Proteins ; biosynthesis ; genetics ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Ki-67 Antigen ; biosynthesis ; Lipoproteins ; biosynthesis ; genetics ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; etiology ; Prostatic Neoplasms ; metabolism ; pathology ; Trans-Activators ; biosynthesis ; genetics

OBJECTIVETo investigate the phosphorylation intensity of MAPK pathway molecular Erk1/2 and the proliferation of prostate cancer cell line PC-3M.

METHODSFlow cytometry and RT-PCR were employed to study the ratio of different cell cycles and phases, respectively, before and after GM-CSF stimulation. Erk1/2 phosphorylation intensity was examined by Western blot simultaneously.

RESULTSThe rate of PC-3M cells at S and G2/M stages and the expression intensity of Ki-67 increased after GM-CSF incubation in a dose-dependent manner. The phosphorylation intensity of Erk1/2 increased remarkably after stimulation with GM-CSF.

CONCLUSIONThe intensification of Erk1/2 phosphorylation is one important molecular mechanism of the proliferation of hormone-independent prostate cancer.

Cell Line, Tumor ; Cell Proliferation ; Granulocyte-Macrophage Colony-Stimulating Factor ; pharmacology ; Humans ; Ki-67 Antigen ; biosynthesis ; Male ; Mitogen-Activated Protein Kinase 1 ; metabolism ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; physiology ; Neoplasms, Hormone-Dependent ; metabolism ; pathology ; physiopathology ; Phosphorylation ; Prostatic Neoplasms ; metabolism ; pathology ; physiopathology ; Signal Transduction ; drug effects

OBJECTIVETo explore the anti-tumor mechanism of norcantharidin (NCTD) for the implanted tumors of human gallbladder carcinoma in nude mice in vivo.

METHODSAnimal model of implanted tumors of human gallbladder carcinoma in nude mice was established. Mice were randomly divided into control, 5-FU, NCTD and NCTD + 5-FU groups and were taken different treatment. The expressions of PCNA, Ki-67, cyclin D1, p27, Bcl-2, Bax, Survivin, nm23/nm23-H1, MMP2 and TIMP2 proteins or genes in each tissue section of every group were determined by immunohistochemistry and RT-PCR.

RESULTS(1) On proliferation-related gene proteins, the expression of PCNA, Ki-67, cyclin D1 was significantly decreased, with significantly increased expression of p27 protein, in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of PCNA mRNA, cyclin D1 mRNA was decreased, with significantly increased expression of p27 mRNA in NCTD group. (2) On apoptosis-related gene proteins, the expression of Bcl-2 was significantly decreased in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of Bcl-2 mRNA, Survivin mRNA was significantly decreased, with significantly increased expression of Bax mRNA in NCTD group. (3) There was significant difference on invasion around tumor and lung metastasis in NCTD group when compared with control group (P < 0.01). On metastasis-related gene proteins, the expression of nm23 and TIMP2 was significantly increased, with significantly decreased expression of MMP2 in paraffin sections of NCTD group when compared with control group (P < 0.05); The expression of nm23-H1 mRNA, TIMP2 mRNA was significantly increased, with significantly decreased expression of MMP2 mRNA in NCTD group.

CONCLUSIONSThe anti-tumor mechanism of NCTD for human gallbladder carcinoma in nude mice might correlated with inhibition of cell proliferation, blockage of cell cycle, induction of cell apoptosis, reducing of cell motility and invasive capability, alteration of the expression of proliferation-, apoptosis- and metastasis-related gene proteins such as PCNA, Ki-67, cyclin D1, p27, Bcl-2, Bax, Survivin, nm23, MMP2 and TIMP2.

Animals ; Apoptosis ; drug effects ; Bridged Bicyclo Compounds, Heterocyclic ; pharmacology ; Cell Proliferation ; drug effects ; Cyclin D1 ; biosynthesis ; genetics ; Gallbladder Neoplasms ; drug therapy ; metabolism ; pathology ; Humans ; Ki-67 Antigen ; biosynthesis ; genetics ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Proliferating Cell Nuclear Antigen ; biosynthesis ; genetics ; RNA, Messenger ; genetics ; bcl-2-Associated X Protein ; biosynthesis ; genetics

OBJECTIVETo investigate the expression of the human mammoglobin (hMAM) mRNA in bone marrow and its clinical significance in the breast cancer patient.

METHODSExpression of hMAM mRNA was detected using nested reverse transcription polymerase chain reaction (RT-PCR) in the bone marrow aspiration sample from 75 breast cancer patients, 15 patients with benign breast lesions and 8 healthy volunteers as control. The possible correlation of hMAM mRNA expression with clinico-pathological parameters and related molecular markers such as Ki67, p53 and VEGF were analyzed.

RESULTSThe sensitivity of RT-PCR in this series reached 10(-6). The hMAM mRNA was found to be positively expressed by RT-PCR in 21 of 75 breast cancer patients with a positive rate of 28.0%. However, hMAM mRNA expression was not detected in the bone marrow aspiration samples from patients with benign breast lesions and healthy volunteers. The hMAM mRNA expression was positively correlated with axillary nodal involvement and progesterone receptor (PR) status (P < 0.05) as well as Ki67 expression in breast cancer tissue (chi2 = 4.936, P = 0.026), but not with age, tumor size, clinical stage, or estrogen receptor (ER) status (P > 0.05).

CONCLUSIONRT-PCR is quite sensitive and has a high specificity in detecting the presence of hMAM mRNA in the bone marrow from breast cancer patients. Thereupon, hMAM mRNA may be useful as a molecular biomarker in detecting disseminated tumor cells (DTC) in the bone marrow of breast cancer patients. Positive hMAM mRNA expression result may have an impact upon therapeutic recommendations and patients' prognostic judgement.

Adult ; Aged ; Biomarkers, Tumor ; genetics ; Bone Marrow ; metabolism ; pathology ; Breast ; metabolism ; pathology ; Breast Neoplasms ; genetics ; pathology ; Breast Neoplasms, Male ; genetics ; pathology ; Carcinoma, Ductal, Breast ; genetics ; pathology ; Female ; Fibroadenoma ; genetics ; pathology ; Humans ; Ki-67 Antigen ; genetics ; Lymphatic Metastasis ; Male ; Mammaglobin A ; Middle Aged ; Neoplasm Proteins ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Receptors, Progesterone ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Uteroglobin ; genetics

To evaluate the expressions of proliferative antigen Ki-67 and apoptosis-antagonizing protein Bcl-2 as well as their clinical significance, immunohistochemistry staining with SAP was used to detect Ki-67 antigen and Bcl-2 protein in 18 cases of children with acute lymphoblastic leukemia (ALL) and 43 cases of adults with ALL. The results showed that the levels of Ki-67 and Bcl-2 expression in children with ALL were lower than that in adults, but only Bcl-2 expression had significant difference. Both in children and in adults, the levels of Ki-67 expression in T-ALL and My(+) ALL were higher than that in B-ALL and null-ALL. The highest complete remission rate (CR) was seen in the group with lower expression of both indexes (Ki-67 and Bcl-2). The lowest CR rate was seen in the group with higher expression of both indexes. It is concluded that the levels of Ki-67 and Bcl-2 expression in children and adults with ALL were closely related with the subtype of ALL and chemotherapeutic effects.
Adolescent ; Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Apoptosis ; physiology ; Child ; Child, Preschool ; Female ; Humans ; Ki-67 Antigen ; biosynthesis ; Male ; Middle Aged ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; drug therapy ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis