Mutation of ING1 gene in laryngeal squamous cell carcinoma and its association with p33ING1b protein expression / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the ING1 gene mutation status in human laryngeal squamous cell carcinoma(LSCC), and the association of p33(ING1b) protein expression with p53 protein expression.@*METHOD@#DNA of LSCC tissue was extracted, and nucleotide of the second exon was amplified and sequenced to determine the chromosome status. The p23(ING1b) and p53 protein expression were detected by immunohistochemistry and the association between them were analyzed.@*RESULT@#No mutation was detected in ING1 gene, but a single polymorphism from GGG to AGG at codon 170 of ING1 gene was found in 2 of the 25 LSCC tissues. The immunohistochemical analysis showed that 4 had positive p33(ING1b) expression. No association was found between p33(ING1b) expression and LSCC clinical features, or between p53 and clinical features. However, significant difference was found between p33(ING1b) and p53 expression. p33(ING1b) tended to be negative in p53 expression positive tissue.@*CONCLUSION@#ING1 gene mutation appears rare in LSCC. In normal physical condition, p33(ING1b) may play a synergistic effect with p53 protein.

Effects of azidothymidine on p33ING1b expression, apoptosis and senescence of TJ905 human glioblastoma cell line / 中华病理学杂志

<p><b>OBJECTIVES</b>To investigate the pharmacological effects of azidothymidine (AZT) on p33ING1b expression, senescence and apoptosis of TJ905 glioblastoma cells.</p><p><b>METHODS</b>TJ905 cells were treated with AZT at a serial concentrations of 50, 100 and 200 µmol/L. Semi-quantitative RT-PCR and cytochemical staining of senescence related-galactosidase (sβ-Gal) were used to evaluate the expression of p33ING1b mRNA and to label the senescent cells at the 1st, 3rd and 6th generations, respectively. In situ cell death detection and single cell gel electrophoresis were used to detect the apoptosis at the 3rd and 6th generations.</p><p><b>RESULTS</b>AZT induced the expression of p33ING1b mRNA and senescence of the tumor cells of the 1st generation in a dosage and time dependent manner. At the 6th generation, the relative amount of p33ING1b RT-PCR product (1.44±0.23) and sβ-Gal labeling index of 200 µmol/L group (45.62±6.74) were significantly higher than those of the 1st (0.95±0.13 and 7.82±2.40) and the 3rd generation cells (1.35±0.23, 26.27±7.17) of the same group, and cells of the same generation in the 50 µmol/L (0.85±0.24, 27.37±6.41) and 100 µmol/L groups (1.23±0.34, 35.49±5.12, P<0.01). There was a significant positive correlation between the p33ING1b mRNA expression and the labeling index of sβ-Gal. Pro-apoptotic effects of AZT became obvious at the 6th generation.</p><p><b>CONCLUSION</b>AZT upregulates the expression of p33ING1b, a possible mechanism in regulating senescence and apoptosis of the TJ905 cells.</p>

Relationship between human papillomavirus 16 infection and the expression of p33(ING1b), human telomerase reverse transcriptase in cervical squamous cell carcinoma of Uygur female in Xinjiang Uygur Autonomous Region / 中国医学科学院学报

<p><b>OBJECTIVE</b>To explore the relationship between human papillomavirus (HPV) 16 infection and the expression of p33(ING1b), human telomerase reverse transcriptase (hTERT) in cervical squamous cell carcinoma of Uygur Female in Xinjiang Uygur Autonomous Region.</p><p><b>METHODS</b>Polymerase chain reaction (PCR) and immunohistochemical methods were used to detect HPV16 infection and the expression of p33(ING1b) and hTERT in the normal control group (n=12), the patients with cervical intraepithelial neoplasm (CIN) (n=34), and the patients with cervical squamous cell carcinoma (SCC) (n=50).</p><p><b>RESULTS</b>In the cervical tissues of Uygur female, the HPV16 infection rate was 0 in control group, 22.2% in the CIN 1 group, 44.0% in CIN 2 & CIN 3 group, and 74.0% in SCC group (P = 0.000). The expression rate of p33(ING1b) decreased was 91.7% in control group, 77.7% in CIN 1 group, 68.0% in CIN 2 & CIN 3 group, and 36.0% in SCC group (P = 0. 000). The expression rate of hTERT was 50.0% in control group, 66.6% in CIN 1 group, 88.0% in CIN 2 & CIN 3 group, and 94.0% in SCC group (P = 0.000). In the cervical tissues of Uygur female, the HPV16 infection rate was negatively correlated with the expression of p33(ING1b) (r = -0.294, P = 0.004), and was positively correlated with the expression of hTERT (r = 0.286, P = 0.005). The expression of p33(ING1b) was negatively correlated with the expression of hTERT (r = -0.361, P = 0.000).</p><p><b>CONCLUSION</b>The infection of HPV 16 correlates with the decreased expression of p33(ING1b) and increased expression of hTERT in the cervical squamous cell carcinoma of Uygur female in Xinjiang.</p>

Expression vector for the inhibitor of growth-1 gene is constructed and the NLS-GFP fusion protein expresses in MRC-5 cells / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To construct the NLS(ING1)-GFP vector, transfer it into MRC-5 cells and establish a cell model expressing NLS (ING1)-GFP fusion protein.</p><p><b>METHODS</b>Firstly, cDNA fragment of nuclear locating sequence (NLS) of inhibitor of growth-1 gene (ING1) was gained by RT-PCR and inserted into multi-clone site of pEGFP-C1 to construct the NLS (ING1)-GFP expression vector. Then the vector was used to transfect the MRC-5 cells to observe the subcellular signal localization of green fluorescence protein (GFP).</p><p><b>RESULTS</b>We successfully constructed the expressing vector of NLS (ING1)-GFP fusion protein. After transferring the fusion expressing vector into MRC-5 cells, we observed that green fluorescence signal located in the cell nucleus. However, the green fluorescence signal located in the cytoplasm in MRC-5 cells transfected with pEGFP-C1 control only expressing GFP.</p><p><b>CONCLUSION</b>In living cells, physiologically p33 ING1b locates absolutely in nucleus. The p33(ING1b) NLS plays a decisive role in the transporting process of subcellular localization.</p>

Expression of P33ING1, P53 and their relationship with apoptosis in anal canal carcinoma / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the expressions of P33ING1, P53 and their relationships with apoptosis in anal canal carcinoma (ACC).</p><p><b>METHODS</b>The expressions of P33ING1, P53 proteins were measured by immunohistochemistry method (SP method), and apoptosis was detected in 42 cases with ACC, 36 cases with anal canal adenoma (ACA) or anal canal papilloma (ACP), and 40 cases with paraanal inflammatory mass(PAIM).</p><p><b>RESULTS</b>The positive expression rates of P33ING1 and P53 proteins were 40.5% (17/42), 97.2% (35/36) and 97.5% (39/40), 50.0% (21/42), 22.2% (8/36) and 27.5% (11/40) respectively, and the average apoptosis indexes(AI) were (10.27+/- 1.23) per thousand, (42.75+/- 0.98) per thousand and (42.67+/- 1.04) per thousand respectively in ACC, ACA or ACP and PAIM. There were significant differences in the positive expression rates of P33ING1, P53 and apoptosis index between ACC and the other two groups respectively (P< 0.05). Among 21 cases of ACC with positive expression of P53 protein,there were 18 cases with P33ING1 negative expression.</p><p><b>CONCLUSIONS</b>P33ING1 expression decrease in ACC, which may play an important role in the carcinogenesis and progression of ACC. P33ING1 and P53 may have an synergistic effect of suppressing cell growth and accelerating cell apoptosis.</p>

Effects of two variants of ING1 expression on tumor cell growth regulation / 中华病理学杂志

<p><b>OBJECTIVE</b>To study effects of alternative transcripts of ING1 transfection on human cancer cell lines.</p><p><b>METHODS</b>p47/ING1A and p33/ING1B expression vehicles were constructed and introduced into a human breast cancer cell line MCF-7 and a human lung cancer cell line PAa, both expressing wild-type p53 protein. Growth characteristics of the transfectants and potentially related genes were analyzed.</p><p><b>RESULTS</b>The levels of p47/ING1A and p33/ING1B protein elevated respectively in tumor cells of MCF-7 and PAa after transfected with p47/ING1A and p33/ING1B, and the latter was much higher than that of the former. Ectopic overexpression of p33/ING1B effectively blocked tumor cell growth and arrested cells in the G(0) approximately G(1) phase of the cell cycle (P < 0.01), while p47/ING1A gave no effect on cell growth or cell cycle. Tumor cells overexpressing p33/ING1B contained more p21(WAF1) protein than that of the control cells, with undisturbed p53 protein level.</p><p><b>CONCLUSIONS</b>Expression of two different transcripts of ING1 may have different effects on tumor cell growth. p33/ING1B may cooperate with p53 in stimulating expression of p21(WAF1) gene, thus to arrest cell cycle and to inhibit tumor cell growth. p33/ING1B may be considered to be a candidate as a partner of p53 in gene therapy.</p>

Study on expression of ING1, human telomerase reverse transcriptase and telomerase-associated protein 1 genes in human gliomas / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the relationship between expressions of ING1 gene and genes of human telomerase reverse transcriptase (hTERT) and telomerase-associated protein 1 (hTP1) in human gliomas.</p><p><b>METHODS</b>The expressions of ING1 mRNA and p33(ING1) protein, hTERT mRNA and protein, and hTP1 mRNA and protein in seventy human glioma specimens with different malignant grades were studied using in situ hybridization and immunohistochemistry.</p><p><b>RESULTS</b>All of the 70 gliomas collected expressed hTP1 mRNA and protein and among them, 62 (88.6%) and 58 (82.9%) out of 70 expressed hTERT mRNA and protein respectively. The quantities of the four kinds of positive cells were correlated positively with one another (r = 0.758 - 0.882, P < 0.000 5), and all of them were significantly fewer in gliomas of WHO grade I - II than in grade III gliomas and the most in grade IV gliomas (P < 0.05 approximately 0.01). 66 (94.3%) and 62 (88.6%) out of 70 gliomas expressed ING1 mRNA and p33(ING1) protein respectively. The quantities of their positive cells were also correlated positively with each other (r = 0.831, P < 0.000 5), but the positive cells were more in gliomas of WHO grade I - II than in grade III gliomas and the fewest in grade IV gliomas (P < 0.01). The quantities of positive cells of ING1 mRNA and p33(ING1) protein were correlated negatively with those of hTERT mRNA and protein as well as hTP1 mRNA and protein respectively (r = -0.211 to -0.384, P < 0.05 approximately 0.001).</p><p><b>CONCLUSIONS</b>The results suggest that all of the parameters concerned are valuable in evaluating the biological behavior of gliomas. In glioma cells, overexpressions of hTERT and hTP1 genes might be significant in inhibiting the expression of ING1 gene. The abnormal expressions of the three genes play possibly the important roles in the development and malignant progression of gliomas.</p>