[Expression of beta-catenin, Glut-1, PTEN proteins in uterine endometrioid adenocarcinoma and its precursor lesions].

OBJECTIVE: To explore the expression of beta-catenin, Glut-1, PTEN in uterine endometrioid adenocarcinoma and their roles in tumorigenesis. METHODS: A total of 83 cases of endometrial hyperplasia were selected and reclassified according to EIN diagnostic criteria. Expressions of beta-catenin, Glut-1 and PTEN proteins were investigated by immunohistochemistry in 10 proliferative endometrium, 83 endometrial hyperplasia and 24 endometrioid adenocarcinoma. RESULTS: (1) 24 EIN (28.9%) lesions were reclassified among 83 previously diagnosed endometrial hyperplasia, of which, 16 of 24 EIN cases (66.7%) had a prior diagnosis of complex atypical hyperplasia. The relation between EIN diagnosis and grade of atypical hyperplasia was not obvious (P > 0.05). (2) Normal (membranous) expression of beta-catenin was present in 10 cases of proliferative endometrium. Abnormal (marked membranous/cytoplasmic, cytoplasmic and/or nuclear or negative) expression rates of beta-catenin in EIN lesions (50%, 12/24) and endometrioid adenocarcinoma (66.7%, 16/24) were significantly higher than that of benign hyperplasia (10.2%, 6/59) respectively (P < 0.01). However, the difference was not significant between EIN lesions and endometrioid adenocarcinomas (P > 0.05). (3) Low level expressions of Glut-1 was present in proliferative endometrium and benign hyperplasia. Overexpression of Glut-1 was present in 58.3% (14/24) of EIN and 70.8% (17/24) of endometrioid adenocarcinoma, respectively, and statistically not significant (P > 0.05). (4) Percentages of loss of PTEN expression showed no difference between EIN lesions (37.5%, 9/24) and proliferative endometrium (2/10), benign hyperplasia (28.8%, 17/59), endometrioid adenocarcinoma (62.5%, 15/24; P > 0.05). However, loss of PTEN expression in endometrioid adenocarcinoma was significantly higher than those in proliferative endometrium and benign hyperplasia (P < 0.05). CONCLUSIONS: Abnormal expression of beta-catenin and overexpression of Glut-1 may be the early events in tumorigenesis of endometrioid adenocarcinoma. The expression of both markers may be useful in distinguishing a benign hyperplasia from EIN and endometrioid adenocarcinoma. Lack of PTEN expression may be the earliest event in endometrial carcinogenesis. However, it can not be used yet as a diagnostic marker for the EIN lesion.

Interleukin-1 gene family polymorphisms from paraffin-embedded archival tissue of Chinese patients with gastric ulcer and gastric cancer.

BACKGROUND/AIMS: Polymorphisms of the interleukin (IL)-1 gene family have been associated with increased risk of gastric ulcer and gastric cancer. The aim of this study was to determine the relationship between single nucleotide polymorphisms (SNPs) in the IL-1 gene family in archival tissue of gastric ulcer and gastric cancer to enable further large scale population investigation in the Chinese Han population from the Wuhan Hubei region. METHODOLOGY: DNA was extracted from archival paraffin-embedded tissue blocks of 53 cases with gastric cancer (22 intestinal type, 29 diffuse type, and 2 mixed type) and 34 cases with gastric ulcer. The tissue specimens had been routinely fixed in unbuffered 10% formalin. Genotyping of SNPs IL-1B T-31C, IL-1B C+3954T and IL-1RN T+2018C was performed using Taqman real-time PCR allelic discrimination technology. RESULTS: The frequency of genotype IL-1B-31C/C seems to be increased in patients with gastric cancer (32.1%) when compared to patients with gastric ulcer (11.8%) (P<0.05). Allele IL-1B+3954T/T was not be found in patients with gastric cancer and gastric ulcer. The quality of genomic DNA extracted from the paraffin-embedded, unbuffered formalinfixed tissue blocks did enable reliable genotyping of all samples. CONCLUSIONS: IL-1B-31C/C may be associated with gastric cancer in patients of Chinese Han population. The genotype of IL-1B3954 T/T may protect against gastric cancer and gastric ulcer in Chinese Han population. Unbuffered formalin-fixed, paraffin-embedded gastric tissue from Chinese patients with gastric ulcer and gastric cancer is a valuable source of DNA, suitable for large scale investigation.

[Clinical analysis of 21 cases of cervical adenosquamous carcinoma].

OBJECTIVE: To study the clinical characteristics, treatment modalities and prognosis of cervical adenosquamous carcinoma. METHODS: The data of 21 patients with adenosquamous cervical cancer who were admitted into Zhongnan Hospital, Wuhan University from Jan 2001 to Dec 2005 were analyzed retrospectively. Six patients received surgical therapy only and 15 patients received combined therapy. RESULTS: Seven cases were with positive pelvic lymph node metastases and 3 cases were with ovarian metastases. The median survival time of the combined therapy group and surgical therapy group was 54 and 20 months, respectively. In stage I and stage II patients, the median survival time of the combined therapy group and surgical therapy group was 66 and 20 months, respectively. The difference was significant between the two groups (P < 0.05). CONCLUSIONS: Combined therapy should be given to patients with adenosquamous carcinoma of the cervix. Surgical therapy and chemotherapy play an important role in the management and prognosis of adenoquamous carcinoma of cervix. Preserve of ovary for patients with adenosquamous carcinoma of the cervix should only be done when the ovary is confirmed free from any malignant involvement by pathology.

Dominant expression of 85-kDa form of cortactin in colorectal cancer.

PURPOSE: Cortactin is commonly expressed in several human cancers, which may alter their invasive or metastatic properties. Eighty five kilodalton form (p85) and 80-kDa form (p80) of cortactin are two separate bands in SDS-PAGE representing different conformational states. The objective of this study was to investigate cortactin expression in colorectal cancer (CRC). EXPERIMENTAL DESIGN: Cortactin expression was studied in an eight paired laser capture microdissection (LCM) CRC tissues and matched non-cancerous epithelia by immunoblotting. The expression in 58 CRC and two cell lines, HCT8 and HCT116, was studied respectively by immunohistochemistry and confocal laser scanning immunofluorescence. RESULTS: Dominant expression of p85 was identified in LCM-procured CRC tissues compared with equal intensity of p85 and p80 forms in non-cancerous tissues, while the amount of total cortactin was approximate. Immunohistochemistry analysis demonstrated that cortactin located in the cytoplasm of tumor cells and adjacent non-cancerous cells, and its expression was negatively correlated with TNM staging and lymphatic invasion status. However, the invasion fronts in 3 of 58 primary tumors and 28 of 39 available lymph node metastases were intensively stained. Further, immunofluorescence analysis showed that cortactin was distributed in cytoplasm and enriched in the front of the extending lamellipodia at adhering side of cultured cancer cells. CONCLUSIONS: Our results demonstrated the dominant expression of p85 form of cortactin in CRC for the first time. The enrichment of cortactin in the invasion front of some tumor cells and in the extending lamellipodia of cultured cancer cells suggests that cortactin may help cancer cell movement.

[Association between expression of survivin and high-risk human papillomavirus infection in cervical cancer and precancerous tissues].

OBJECTIVE: To investigate the expression of survivin and its association with high-risk human papillomavirus (HPV) infection in cervical cancer and precancerous tissues. METHODS: In 76 patients with cervical intraepithelial neoplasia (CIN) or cervical carcinoma, the expression of survivin was detected by immunohistochemistry and the infection rates of high-risk HPV including HPV16, 18, 31, 33, 52, and 58 were assayed by touch-down PCR, with 10 cases of normal cervical specimens serving as the control. RESULTS: Survivin expression tended to increase along with the tumor progression (P<0.05), and was associated with the histological grade of invasive carcinoma (P<0.05) but not with the patients' age, clinical stage or histological classification (P>0.05). The positivity rate of high-risk HPV increased only in association with tumor progression (P<0.05) but not with the patients' age, clinical stage or histological classification (P>0.05). Positive correlation between the expression of survivin and high-risk HPV infection was observed in cervical carcinoma. CONCLUSION: Survivin might play an important role in the occurrence and development of cervical carcinoma in coordination with high-risk HPV.

Syndecan-1 and E-cadherin expression in differentiated type of early gastric cancer.

AIM: To elucidate the role and alterations of syndecan-1 and E-cadherin expression in different cellular phenotypes of differentiated-type gastric cancers (DGCs). METHODS: A total of 120 DGCs at an early stage, and their adjacent mucosa, were studied both by immunohistochemistry. Syndecan-1 and E-cadherin were assessed by immunohistochemical staining with anti-syndecan-1 and anti-E-cadherin antibodies, respectively. Based on immunohistochemistry, DGCs and their surrounding mucosa were divided into four types: gastric type (G-type), ordinary type (O-type), complete-intestinal type (CI-type), and null type (N-type). RESULTS: Syndecan-1 expression was significantly lower in G-type cancers (29.4%) than in O-type (79.6%) and CI-type cancers (90%) (P<0.05, respectively), but E-cadherin did not show this result. In addition, syndecan-1 expression was significantly reduced in DGCs comprised partly of poorly differentiated adenocarcinoma or signet-ring cell carcinoma, compared to DGCs demonstrating papillary and/or tubular adenocarcinoma (P<0.05). G-type intestinal metaplasia (IM) surrounding the tumors was observed in 23.8% of G-type, 4.9% of O-type, and 6.7% of CI-type cancers (P<0.05; G-type vs O-type). Reduction of syndecan-1 expression was significant in G-type IM (25%) compared to non-G-type IM (75%; P<0.05). CONCLUSION: Loss of syndecan-1 plays a role in the growth of G-type cancers of DGCs at an early stage, and the reduction of syndecan-1 expression in IM surrounding the tumors may influence the growth of G-type cancer.

[Relationships among expressions of hTERT, MDR1, MRP mRNA, and C-myc protein in non-small cell lung cancer].

BACKGROUND & OBJECTIVE: The latest researches showed that myc protein could up-regulate the expression of human telomerase reverse transcriptase (hTERT), multidrug resistance gene 1(MDR1), multidrug resistance-related protein (MRP) in some kinds of tumors, and hTERT is correlated with efficiency of anti-tumor chemotherapy. This study was to investigate relations among expressions of hTERT, MDR1, MRP mRNA, and C-myc protein in non-small cell lung cancer (NSCLC). METHODS: Expressions of hTERT, MDR1, MRP mRNA in 113 cases of NSCLC tissues were detected by in situ hybridization, expression of C-myc protein was detected by SP immunohistochemistry, their correlations with clinicopathologic features of NSCLC were statistically analyzed. RESULTS: Positive rates of hTERT, MDR1, MRP mRNA, and C-myc protein in NSCLC tissues were 80.5%, 51.3%, 80.5% and 68.1%, respectively. Expressions of MDR1, MRP mRNA, and C-myc protein were significantly related to that of hTERT mRNA (P<0.05). Expression of C-myc protein did not correlate with expression of MDR1 or MRP mRNA. All 4 factors have no correlation with clinicopathologic features of NSCLC (P>0.05). CONCLUSION: Expression of hTERT mRNA may be related to those of MDR1, MRP mRNA, and C-myc in NSCLC. Overexpression of C-myc protein may be one of the molecular regulatory mechanisms of hTERT mRNA.

[Expression of NFkappaB p65 and its target genes in gastric cancer and precancerous lesions].

OBJECTIVE: To study the expression of NFkappaB p65 and its target genes in intestinal metaplasia (IM), dysplasia (Dys), gastric cancer (GC) infected with Helicobacter pylori (Hp) and explore the mechanism of infection by cytotoxin-associated antigen A expressing Hp (CagA(+)Hp) in the development of gastric cancer. METHODS: CagA antibody in blood sample of 289 patients was determined by ELISA. Hp was detected by rapid urease test and Warthin starry staining. Expression of NFkappaB p65 and its target genes in IM, Dys and GC was examined by immunohistochemistry. RESULTS: In IMI approximately II, IMIII, DysI, DysII approximately III and GC, the expression of NFkappaB p65 was significantly higher in patients with CagA(+)Hp infection than those without CagA Hp infection. In IMIII and DysII approximately III, the expression of NFkappaB p65, c-myc, CyclinD(1) and bcl-xl was significantly higher in patients with CagA Hp infection than those without CagA Hp infection. In gastric cancer infected with CagA(+)Hp, the expression of NFkappaB p65, c-myc, CyclinD(1) and bcl-xl was significantly higher in intestinal type than in diffuse type. CONCLUSION: There are different mechanisms in intestinal type and diffuse type in the development of gastric cancer. The occurrence of intestinal type gastric cancer is associated with CagA(+)Hp infection which by NFkappaB p65 upregulating the expression of c-myc, CyclinD(1),bcl-xl in patients with IMIII, DysII approximately III. It may be an effective method to prevent gastric cancer by inhibiting NFkappaB p65.

[Expression and significance of telomerase reverse transcriptase and its regulators in non-small cell lung carcinoma].

BACKGROUND & OBJECTIVE: Telomerase is a ribonucleoprotein complex, which is silent in normal human somatic cells but may be reactivated by a series of regulators, and cause tumorigenesis. As a critical factor of telomerase activity, much progression has been achieved in the study of regulation of human telomerase reverse transcriptase, but the detail mechanism is still not clear. This study was designed to investigate the relationship of expression of human telomerase reverse transcriptase mRNA (hTERT mRNA) and its regulators including c-myc, mutant p53, protein kinase C alpha (PKCalpha) with clinicopathological significance of expression of the four markers in non-small cell lung carcinoma (NSCLC). METHODS: The expression of hTERT mRNA in 113 NSCLC specimens were detected by in situ hybridization, and the expression of c-myc, mutant p53, and PKCalpha in the same specimens were detected by immunohistochemistry. RESULTS: The positive rates of 113 NSCLC samples for hTERT mRNA, c-myc, mutant p53, and PKCalpha were 80.5%, 68.1%, 61.9%, and 85.0%, respectively. The differences were statistically significant among the expression of hTERT mRNA, c-myc, and PKCalpha (P< 0.05 or P< 0.01), but was not significant between the expression of mutant p53 and hTERT mRNA, c-myc as well as PKCalpha. The expression of mutant p53 was associated with carcinoma cell differentiation (P< 0.05), and its positive rate gradually increased with the extend of differentiation of carcinoma cell. The expression of hTERT mRNA, c-myc, and PKCalpha were not associated with carcinoma cell differentiation. There was no relationship of expression of the four markers with histological types, TNM stages, and lymph node metastasis status. CONCLUSION: C-myc and PKCalphawere associated with the expression of telomerase.

Expression of nuclear factor-kappa B and target genes in gastric precancerous lesions and adenocarcinoma: association with Helicobactor pylori cagA (+) infection.

AIM: To examine the expression of nuclear factor kappaB (NF-kappaB) and its target genes in intestinal metaplasia (IM), dysplasia (DYS) and gastric carcinoma (GC) infected with Helicobacter pylori (H pylori) and to investigate the mechanism underlying H pylori cytotoxin associated gene A (cag A) infection leading to gastric adenocarcinoma. METHODS: Expressions of NF-kappaB/p65 and its target genes: c-myc, cyclinD1 and bcl-xl were immunohistochemically examined in 289 cases of gastric biopsy and resection specimens from patients with IM, DYS and GC infected with H pylori. H pylori in the above mentioned tissues was detected by Warthin-Starry stain and rapid urease tests. IgG antibody to cagA in sera of the patients was measured by ELISA. RESULTS: The positive rates of NF-kappaB/p65 were significantly higher in groups with cagA of IMI-II(28/33), IM III(48/52), DYSI(27/31), DYS II-III(28/32), GC(35/40) than in groups without cagA of IMI-II(4/17), IMIII(3/20), DYSI(3/20), DYSII-III(6/21), GC(10/23). The expressions of c-myc, cyclinD1, and bcl-xl were significantly higher in groups with cagA of IM III(47/52, 49/52, 46/52), DYSII-III(29/32, 26/32, 25/32) than in groups without cagA of IM III(8/20, 7/20, 5/20), DYSII-III(10/21, 8/21, 3/21), which were in conformity with the expression of NF-kappaB in IM III, and DYSII-III. A significantly higher expression level of NF-kappaB/p65, c-myc, cyclinD1 and bcl-xl was detected in intestinal type GC(27/28, 18/28, 22/28, 24/28) than in diffuse type GC(8/12, 3/12, 3/12, 6/12), respectively. CONCLUSION: There may be two different molecular mechanisms in the occurrence of intestinal and diffuse type gastric carcinomas. Intestinal type gastric carcinoma is strongly associated with high expression of c-myc, cyclinD1 and bcl-xl through NF-kappaB/p65 activated by H pylori cagA. Inhibiting the activity of NF-kappaB is an effective and promising way to prevent intestinal type gastric carcinoma.

[Relationship between Helicobacter pylori infection and expression of c-myc, Bcl-2, and Bax protein in different gastric mucosa lesions].

BACKGROUND & OBJECTIVE: Helicobacter pylori (HP) has been believed to be a carcinogen of gastric carcinoma. However, its mechanism was yet not clearly understood. This study was designed to investigate the relationship between HP infection and gastric epithelial cell proliferation as well as apoptosis in different gastric mucosa lesions and elucidate the probable mechanism of gastric carcinogenesis relating with HP infection. METHODS: A total of 272 cases were available for the study including 42 cases of chronic gastritis (CG), 46 cases of intestinal metaplasia I or II (IM I- II), 25 cases of intestinal metaplasia III (IM III), 21 cases of mild dysplasia (Dys I), 54 cases of modest or severe dysplasia (Dys II- III), and 84 cases of gastric cancer (GC). HP infection was detected by Warthin-Starry bacterium staining method and streptavidin-peroxidase (SP) immunohistochemical method. HID-AB(pH2.5)- PAS method was used to define the quality of mucus. The expression of c-myc, Bcl-2, and Bax were detected using SP immunohistochemical method. The chi-square test and the Fisher's exact probability test were used to compare the frequencies. RESULTS: (1)The expression of c-myc and Bcl-2 increased as gastric mucosa lesions developed from CG,IM,Dys to GC,but the expression of Bax decreased. The expression of c-myc was significantly higher in GC than that in Dys II- III and IM III(all P< 0.01), but the expression of Bax was significantly lower in GC than that in Dys II- III and IM III(P< 0.05 or P< 0.01). (2)The expression of c-myc in IM III and Dys II- III with HP infection was 62.50% and 66.67%,respectively, significantly higher than that without infection(11.11%,27.78%,all P< 0.05). The expression of Bax in CG, IM I- II and IM III with HP infection were 87.10%, 81.25%, and 62.50%, respectively, significantly higher than those without infection (54.55%, 42.86%, 11.11%, all P< 0.05). Furthermore HP infection was associated with the expression of Bcl-2 in IM III, Dys II- III and GC (P< 0.05 or P< 0.01). CONCLUSION: HP infection can cause serious imbalance between cell proliferation and apoptosis in the precancerous lesions (IM III and Dys II- III), giving chances for gastric carcinogenesis.

[Multidrug resistance and its relationship with neuroendocrine differentiation in non-small cell lung carcinoma].

OBJECTIVE: To study the multidrug resistance (MDR) of non-small lung cancer (NSCLC) and its relationship with neuroendocrine (NE) differentiation. METHODS: NSCLC samples from 113 untreated patients were analyzed immunohistochemically with antibodies to glutathion-s-transferase-pi (GST-pi), multidrug resistance associated protein (MRP), lung resistance associated protein (LRP), neuro-specific enolase (NSE), synaptophysin (SYN) and chromogranin (CgA). RESULTS: (1) The expression of the three proteins was significantly associated with the type of lung carcinoma (P < 0.05), but not with the differentiation and lymph node metastasis. The expression of GST-pi was significantly related with MRP, MRP and LRP (P < 0.05). (2) The positive rates of the NE markers were: NSE, 53.1%; SYN, 26.6%; CgA, 6.2%; and 21.2% for at least two markers. The expression of at least 2 markers was associated with the degree of differentiation (P < 0.05), but not with the type of lung cancer and lymph node metastasis. (3) The expression of the three multidrug resistance related proteins in the positive group for at least 2 markers was significantly lower than that in the negative group (P < 0.05). CONCLUSIONS: The over-expressions of GST-pi, MRP and LRP are important causes of primary multidrug resistance in NSCLC. The differentiation of NE may be one of the factors involved in multidrug resistance.

Helicobacter pylori infection generated gastric cancer through p53-Rb tumor-suppressor system mutation and telomerase reactivation.

AIM: To investigate the relationship between Helicobacter pylori (H.pylori) infection and the expressions of the p53, Rb, c-myc, bcl-2 and hTERT mRNA in a series of diseases from chronic gastritis (CG), intestinal metaplasia type I or II(IMI-II), intestinal metaplasia type III (IMIII), mild or modest dysplasia (DysI-II), severe dysplasia (DysIII) to gastric cancer(GC) and to elucidate the mechanism of gastric carcinogenesis relating to H.pylori infection. METHODS: 272 cases between 1998 and 2001 were available for the study including 42 cases of CG, 46 cases of IMI-II, 25 cases of IMIII, 48 cases of DysI-II, 27 cases of DysIII, 84 cases of GC. H.pylori infection and the expressions of p53, Rb, c-myc, bcl-2 were detected by means of streptavidin-peroxidase (SP) immunohistochemical method. HTERT mRNA was detected by in situ hybridization (ISH). RESULTS: The expressions of p53, Rb, c-myc, hTERT mRNA and bcl-2 were higher in the GC than in CG, IM, Dys. The expression of c-myc was higher in IMIII with H.pylori infection (10/16) than that without infection (1/9) and the positive rate in DysI-II and DysIII with H.pylori infection was 18/30 and 13/17, respectively, higher than that without infection (4/18 and 3/10, respectively). In our experiment mutated p53 had no association with H.pylori infection, the expression of Rb was associated with H.pylori infection in GC, but the p53-Rb tumor-suppressor system abnormal in DysI-II cases, DysIII and GC cases with H.pylori infection was 21/30, 15/17 and 48/48 respectively, higher than non-infection groups (4/18, 3/10, 28/36). Furthermore the level of hTERT mRNA in GC with H.pylori infection (47/48) was higher than that without infection (30/36), however the relationship between bcl-2 and H.pylori was only in IMIII. C-myc had a close association with hTERT mRNA in DysIII and GC (P=0.0 253,0.0 305 respectively). CONCLUSION: In the gastric carcinogenesis, H.pylori might cause the severe imbalance of proliferation and apoptosis in the precancerous lesions (IMIII and GysIII) first, leading to p53-Rb tumor-suppressor system mutation and telomerase reactivation, and finally causes gastric cancer.

[Relationship between Helicobacter pylori infection and expressions of tumor suppressor genes in gastric carcinoma and related lesions].

BACKGROUND & OBJECTIVE: Helicobacter pylori (Hp) are believed to be a carcinogen of gastric carcinoma. However, its mechanism was yet not clearly understood. p53, p21WAF1, and p16 are main negative regulator genes of cell cycle. This study was designed to investigate the relationship between these 3 genes and Hp infection. METHODS: The authors examined the expression of these 3 tumor suppressor genes and Hp infection in 65 cases with chronic atrophic gastritis (CAG), 93 cases with intestinal metaplasia(IM), 94 cases with gastric epithelial dysplasia (GED) and 60 cases with gastric carcinoma (GC) using HID-AB (pH 2.5)-PAS, SP immunohistochemistry staining, and Warthin-Starry staining. RESULTS: For CAC, IM stage I-II, IM stage III, GED stage I, GED stage II-III, and GC, the positive expression rates of p53 were 0, 1.64%, 6.25%, 5.45%, 23.08%, and 70.00%, respectively (increased with pathological process); the positive expression rates of p21WAF1 were 100%, 95.08%, 100%, 100%, 71.79%, and 45.00%, respectively; the positive expression rates of p16 were 83.08%, 81.97%, 78.13%, 89.09%, 69.23%, and 40.00%, respectively. All the expression of these 3 genes showed significant difference between GED stage II-III and GED stage I, GC and GED stage II-III. In the same pathological changes, the positive expression rate of these 3 genes was higher in Hp infection group than in no Hp infection group, while there was no significant difference (P > 0.05). CONCLUSION: The mutation of p53 and inactivation of p21WAF1 and p16 play an important role in carcinogenesis of stomach. However, Hp infection was not associated with the abnormal expression of these 3 genes.