Geranylgeranylacetone promotes human osteosarcoma cell apoptosis by inducing the degradation of PRMT1 through the E3 ubiquitin ligase CHIP.

Geranylgeranylacetone (GGA), an inducer of heat shock proteins, exerts anticancer activity in some tumours. However, the effect of GGA on human osteosarcoma (OS) has not been reported. This work is designed to evaluate the effect of GGA on the proliferation and apoptosis of human OS cells and to explore the underlying mechanisms. It was found that GGA markedly inhibited the proliferation and induced apoptosis of U-2 OS cells in a dose-dependent manner and also up-regulated the expression of heat shock protein 70 (Hsp70). The degradation and ubiquitination of protein arginine N-methyltransferase 1 (PRMT1) were obviously enhanced in U-2 OS cells with CHIP overexpression and GGA treatment. The expression of PRMT1 was reversed in GGA-treated cell after CHIP knockdown. The turnover of PRMT1 was obviously faster in cells overexpressing CHIP than that in control cells. The methylation and activity of STAT3 were induced by PRMT1, resulting in the inhibition of FAS transcription. Overexpression of PRMT1 reversed the effect of GGA on activation of apoptosis-related proteins and U-2 OS cell apoptosis. The expressions of PRMT1 were significantly up-regulated in OS tissues compared with the adjacent normal tissues and benign bone tumours. In conclusion, GGA promotes the degradation of PRMT1 through the Hsp70-CHIP-mediated proteasome pathway, thereby inducing the FAS-triggered cell apoptosis. Inhibition of PRMT1 may be a potential therapeutic strategy for OS patients.

IMP3 Predicts Invasion and Prognosis in Human Lung Adenocarcinoma.

PURPOSE: Insulin-like growth factor II mRNA-binding protein 3 (IMP3) is an oncofetal protein associated with several aggressive and advanced cancers. Whether IMP3 can predict invasion, and prognosis in patients with human lung adenocarcinoma (LAC) remains unclear. METHODS: Ninety-five LAC and 75 non-tumor lung tissue samples were included in a tissue microarray. IMP3 expression was assessed by immunohistochemical examination. Correlation between IMP3 expression levels, clinicopathological characteristics, and overall prognosis was evaluated. In a separate in vitro study, RNA interference method was applied for knockdown of IMP3 gene in human LAC cell lines. Invasive potential of LAC cells was then evaluated by transwell migration assay. RESULTS: IMP3 immunoreactivity was observed in 39 out of 95 (41.1 %) LAC patients, but not in non-tumor lung tissues. IMP3 expression levels were closely associated with histological grade (P = 0.037), TNM stage (P = 0.034), and lymph node metastasis (P = 0.011). Patients presenting with positive IMP3 expression (P = 0.000), an advanced TNM stage (P = 0.000), and lymph node metastasis (P = 0.001) had a worse overall survival, compared to those lacking these characteristics. Both IMP3 expression (hazard ratio [HR], 2.310; 95 % confidence interval [CI] 1.192-4.476; P = 0.013) and TNM stage (HR 2.338; 95 % CI 1.393-3.925; P = 0.001) were independent predictors of poor prognosis. The invasive potential of LAC cells was significantly inhibited by IMP3 knockdown. CONCLUSION: IMP3 appears to play an important role in tumor invasion in patients with LAC and may serve as a useful prognostic biomarker in these patients.

Candida albicans and bacterial vaginosis can coexist on Pap smears.

OBJECTIVE: The aim of our study was to study the relationship of blastospores and pseudohyphae in Papanicolaou (Pap) smears and Nugent scores for bacterial vaginosis (BV). STUDY DESIGN: A total of 471 Pap smears with Candida albicans were reviewed. The presence of blastospores and pseudohyphae was established. The Pap smears were restained with the Gram stain method to evaluate the bacterial flora according to the Nugent scoring system. RESULTS: Of the 471 Pap smears, blastospores and pseudohyphae were observed in 62.8% (296/471) and 37.2% (175/471) of the smears, and displayed symptoms in 4.4% (13/296) and 43.4% (76/175), respectively. A significant difference was found between these 2 groups (p < 0.0001). A positive BV Nugent score (≥ 7) was found in 22.1% (104/471) of the C. albicans cases. Blastospores and pseudohyphae with BV were 14.2% (42/296) and 35.4% (62/175), respectively. These high Nugent scores indicate statistically significant differences (p < 0.0001). CONCLUSION: C. albicans and BV can coexist. The presence of blastospores in these C. albicans cases was negatively related to symptoms.

Decreased Expression of Karyopherin-α 1 is Related to the Malignant Degree of Cervical Cancer and is Critical for the Proliferation of Hela Cells.

Karyopherin α (KPNA) proteins are involved in nucleocytoplasmic trafficking and are critical for protein subcellular localization. Recent studies have suggested that KPNA proteins are abnormally expressed in various solid tumors. The objective of this study was to investigate the expression of KPNA1 and KPNA2 in cervical cancer tissue with different histologic grades and cell lines, as well as the effects of the KPNA1 expression level on Hela cell proliferation. We collected the medical data of 106 patients with cervical cancer and investigated the protein expression of KPNA1 and KPNA2 by immunohistochemistry and western blot. The results revealed a significantly lower expression of KPNA1 in cervical cancer compared to normal tissue. Conversely, stronger staining intensity for KPNA2 was observed in cervical tumor samples. The expression levels of KPNA1 and KPNA2 were significantly associated with the tumor histologic grade. The weakest KPNA1 expression and strongest staining for KPNA2 were observed in grade III tumor tissue. The expression levels of KPNA1 were lower in Hela and C33A cells compared with normal human cervical epithelial cells; however, the expression of KPNA2 exhibited an opposite trend. The up-regulation of KPNA1 significantly suppressed the proliferation of Hela cells and relevant proteins expression, as well as promoted transportation of IRF3 into nucleus. Our results suggest the downregulation of KPNA1 expression is related to the malignant degree of cervical cancer and is closely associated with the proliferation of cervical cancer cells.

Morphological features of cell blocks prepared from residual liqui-PREP samples can distinguish between high-grade squamous intraepithelial lesions and squamous cell carcinomas.

OBJECTIVE: To evaluate the diagnostic value and compare morphological features of cell block sections of high-grade squamous intraepithelial lesions (HSIL) and squamous cell carcinomas (SCC). STUDY DESIGN: A total of 135 cell blocks were prepared from residual Liqui-PREP samples. Of these, 43 biopsy-confirmed cases (24 HSIL and 19 SCC) were reviewed. Morphological features determined included cell clusters, epithelial-stromal interface, stromal invasion and tumor necrosis. RESULTS: Ninety-three percent (40/43) of cell block diagnoses were consistent with histological diagnoses, which was better than the cytological diagnoses (81.4%; 35/43). The mean cell block size was 0.54 cm (range, 0.3-1.0 cm) for HSIL and 0.84 cm (range, 0.4-1.4 cm) for SCC (p < 0.0001). Cell clusters were present in 70.8% (17/24) of HSIL and 100% (19/19) of SCC (p < 0.0001). No epithelial-stromal interface, stromal invasion or tumor necrosis was observed on HSIL cell block sections, which is in contrast to the 84.2% (16/19), 68.4% (13/19) and 42.1% (8/19) on SCC cell blocks, respectively (p < 0.05). CONCLUSION: Cell blocks may increase the diagnostic accuracy of liquid-based cytology. The presence of stromal invasion, epithelial-stromal interface and tumor necrosis on cell block sections may be useful for accurate SCC diagnosis.

KPNA2 expression is a potential marker for differential diagnosis between osteosarcomas and other malignant bone tumor mimics.

BACKGROUND: Karyopherin α2 (KPNA2), a member of the karyopherin α family, has been studied in several cancers but has not yet been substantially investigated in malignant bone tumors. The purpose of the current study was to evaluate the KPNA2 expression level and its utility as a novel diagnostic biomarker in osteosarcomas and malignant bone tumor mimics, such as chondrosarcomas and Ewing sarcomas (ESs). METHOD: We investigated the expression of KPNA2 protein by immunohistochemistry on paraffin-embedded surgical specimens from 223 patients with malignant and benign bone tumors, including 81 osteosarcomas, 42 chondrosarcomas, 15 ESs, 28 osteoid osteomas, 20 osteochondromas and 37 chondroblastomas. Immunoreactivity was scored semiquantitatively based on staining extent and intensity. RESULTS: Sixty-seven of 81 (82.7%) osteosarcoma, zero of 42 (0%) chondrosarcoma and one of 15 (6.7%) ES samples showed immunoreactivity for KPNA2. Negative KPNA2 expression was observed in all benign bone tumors. The expression of KPNA2 in osteosarcoma samples was much higher than that in chondrosarcoma and ES samples (P < 0.001). The sensitivity and specificity of KPNA2 immunoexpression for detecting osteosarcoma were 82.7 and 100%, respectively. Several subtypes of osteosarcoma were analyzed, and immunostaining of KPNA2 was frequent in osteoblastic samples (90.9%), with 39 samples (70.9%) showing strong-intensity staining. KPNA2 positivity was observed in ten of 13 (76.9%) chondroblastic, two of 6 (33.3%) fibroblastic, three of 4 (75%) telangiectatic and two of 3 (66.7%) giant cell-rich osteosarcoma samples. The strongest intensity staining was observed in osteoblastic osteosarcoma. CONCLUSION: KPNA2 is frequently expressed in osteosarcomas, particularly in osteoblastic and chondroblastic tumors, but is rarely positive in chondrosarcomas and ESs. This feature may aid in distinguishing between osteosarcoma and other bone sarcoma mimics. This report supports KPNA2 as a novel marker for the diagnosis of osteosarcoma.

Analysis of IMP3 expression in primary tumor and stromal cells in patients with colorectal cancer.

Insulin-like growth factor II mRNA-binding protein 3 (IMP3) is an oncofetal protein upregulated in tumor cells during carcinogenesis. The aim of the present study was to investigate the expression status of IMP3 in colorectal cancer (CRC) tissues and its clinical significance. Immunostaining was performed in 130 CRC samples, the association of IMP3 expression with clinicopathological characteristics was assessed and 58 patients were selected for survival analysis. To the best of our knowledge, the present study describes for the first time the expression of IMP3 in tumor stromal components of CRC. Stromal expression of IMP3 was detected in 24/130 (18.5%) CRC tissue specimens and was associated with tumor-node-metastasis (TNM) stage (stage III-IV, P=0.003), lymph node metastasis (P=0.006), lympho-vascular invasion (P=0.003), tumor border (P=0.013). Tumoral expression of IMP3 was detected in 94/130 (72.3%) of CRC specimens and was associated with T classification (T3-T4, P=0.027), tumor-node-metastasis (TNM) stage (stage III-IV, P=0.011), lymph node metastasis (P=0.048), tumor budding (>10 buds, P=0.005). Further study indicated that patients with IMP3 expressed in tumor cells and tumor stroma tend to have poorer overall survival rates (P=0.02 and P=0.06, respectively). Moreover, tumoral expression of IMP3 and TNM stage were identified to be independent prognostic factors in CRC. IMP3 was not only expressed in tumor cells but also in stroma cells. Stromal expression of IMP3 was associated with lymph node metastasis and advanced tumor TNM stage. Moreover, the survival analysis indicated that there is a significant association between IMP3 expression in tumor cells and a poorer overall survival rate in patients with CRC. The expression of IMP3 maybe a predicted factor for CRC patient.

IMP3 expression in biopsy specimens as a diagnostic biomarker for colorectal cancer.

No single biological marker is used in routine diagnosis of colorectal cancer (CRC) in endoscopic biopsies. IMP3 is a good independent prognostic biomarker for CRC. However, the expression of IMP3 in hyperplastic polyp (HP) and adenoma has not yet been studied. Moreover, no studies have established the diagnostic value of IMP3 in biopsies. This study aims to assess IMP3 expression in HP, adenoma, and CRC in resection specimens and to investigate its value in diagnosis of CRC in biopsies. A total of 1328 specimens (633 of polypectomy, 395 surgical resections, 300 biopsies) were retrospectively analyzed. IMP3 expression was observed in 0 of 197 (0%) normal tissues, 0 of 130 (0%) HPs, 14 of 504 (2.8%) adenomas, and 139 of 197 (70.6%) CRCs. IMP3 was found to be overexpressed in CRC compared with adenoma (P<.001). Among the 300 biopsies, 56 were diagnosed as adenoma, and 244 were CRCs. Of the 56 adenoma cases, 22 (39.3%) were confirmed, whereas 34 (60.7%) were diagnosed as CRC in resection specimens. All 244 CRC biopsies were confirmed by resection specimens. IMP3-positive expression was observed in 204 of 300 (68.0%) biopsies, including in 22 of 56 (39.3%) adenomas and 182 of 244 (74.6%) CRCs. All IMP3-positive expressions in the biopsies were finally diagnosed as CRC. Our findings demonstrated that IMP3 is a reliable marker for the diagnosis of CRC in endoscopic biopsies.

Comparison of Uterine Receptivity between Fertile and Unexplained Infertile Women by Assessment of Endometrial and Subendometrial Perfusion Using Contrast-Enhanced Ultrasound: Which Index is Better--Peak Intensity or Area under the Curve?

The goal of this study was to compare uterine receptivity between women with normal fertility and those with unexplained infertility during natural cycles by assessment of endometrial and subendometrial perfusion using contrast-enhanced ultrasound (CEUS). We wanted to determine the better index: peak intensity (PI) or area under the curve (AUC). Thirty women with unexplained infertility were recruited into the study group, and 30 women with normal fertility were recruited into the control group. All women underwent CEUS during the late proliferative phase, ovulation phase, and implantation window of a menstrual cycle. Endometrial PI, endometrial AUC, subendometrial PI and subendometrial AUC were analyzed. In the late proliferative phase, the control group had a significantly higher endometrial PI (p < 0.001) as well as subendometrial PI (p < 0.001) and AUC (p = 0.004) than the study group. In the ovulation phase, the control group had a significantly higher endometrial PI (p < 0.001) and AUC (p = 0.021), as well as subendometrial PI (p < 0.001) and AUC (p = 0.003). During the implantation window, there were no significant differences between the two groups. Only subendometrial PI underwent a significant periodic change during the menstrual cycle in both groups. This finding was further confirmed by evaluation of the microvessel density of endometria. In conclusion, CEUS can be used to assess endometrial and subendometrial perfusion to evaluate uterine receptivity. Subendometrial PI was the most sensitive index compared with endometrial PI, endometrial AUC and subendometrial AUC.

IMP3 expression in biopsy specimens of colorectal cancer predicts lymph node metastasis and TNM stage.

IMP3 is associated with lymph node metastasis and TNM stage and is a good independent prognostic biomarker for colorectal cancer (CRC). However, the expression status and clinical implication of IMP3 in biopsy specimens have not yet been studied. We aim to address whether the presence of IMP3 expression in preoperative biopsies of CRC could predict lymph node metastasis and TNM stage. In this study, we examined IMP3 expression in paired biopsy and resection specimens of 71 CRC and analyzed the correlation of IMP3 expression with clinicopathological parameters. In the biopsy specimens, IMP3 positive expression was observed in 56 of 71 cases (78.9%) whereas negative expression was observed in 15 of 71 cases (21.1%). In the resection specimens, IMP3 positive expression was detected in 83.1% cases (59/71) whereas negative expression was detected in 16.9% cases (12/71). The absolute concordance rate between biopsy and resection specimens was 90.1% (64/71). The Spearman correlation test documented the existence of a strong linear correlation between the percentage of IMP3-positive cells in the biopsy and resection specimen (r = 0.629; P < 0.001). IMP3 expression in resection specimens was significantly related to histological grade (P = 0.043), T classification (P = 0.035), lymph node metastasis (P = 0.023), TNM stage (P = 0.007), tumor border (P = 0.049) and tumor budding (P = 0.012). IMP3 expression in biopsy specimens was significantly related to lymph node metastasis (P = 0.004), TNM stage (P = 0.005) and tumor budding (P = 0.001). In conclusion, IMP3 expression in biopsy specimens could be used to predict lymph node metastasis and TNM stage in CRC patients.

[Value of serum IgA/C3 ratio in the diagnosis of IgA nephropathy and its correlation with the clinicopathological features].

OBJECTIVE: To investigate the value of serum IgA/C3 ratio in the diagnosis of IgA nephropathy and explore its relationship with the clinicopathological features of the patients. METHODS: Sixty-six patients with IgA nephropathy, 111 with other glomerular diseases, and 40 healthy control subjects without kidney disease were tested for serum IgA and C3 levels using CRM470 adjusted standardized immune turbidimetric method, and the IgA/C3 ratio was calculated. According to Oxford and Lee's classification criteria, we analyzed the pathological grades of the renal biopsy samples from patients with IgA nephropathy. The ROC curve was used to assess the value of serum IgA and IgA/C3 ratio in predicting IgA nephropathy. RESULTS: Patients with IgA nephropathy had an elevated serum IgA/C3 ratio than those with other glomerular diseases and the control subjects, with an area under the ROC curve of 0.776. An elevated serum IgA/C3 ratio was not found to significantly correlate with the pathological grade of renal biopsy samples in patients with IgA nephropathy. CONCLUSION: In the absence of renal biopsy findings, serum IgA/C3 ratio can help in the diagnosis of IgA nephropathy.

A fine decision tree consisted of CK5/6, IMP3 and TTF1 for cytological diagnosis among reactive mesothelial cells, metastatic adenocarcinoma of lung and non-lung origin in pleural effusion.

The utility of combination with CK5/6, IMP3 and TTF1 to differentiate among reactive mesothelial cells (RMs), metastatic adenocarcinoma of lung (LAC) and non-lung (NLAC) origin was investigated by using immunocytochemistry (ICC) and conventional PCR (C-PCR) in pleural effusion. A total of 108 cell blocks (32 RMs, 51 LAC and 25 NLAC were evaluated by ICC for CK5/6, IMP3 and TTF1 protein expression. In addition, we further performed C-PCR for amplification of CK5/6, IMP3 and TTF1 DNA from 28 specimens (9 MAC and 7 RMs, 6 LAC and 6 NLAC) for molecular diagnosis. CK5/6 staining was observed in the majority of reactive specimens (78.1%) and was rare in adenocarcinoma cells (14.5%), whereas the opposite was true for IMP3 and TTF1. We found a high frequency of TTF1 positivity (76.5%) in LAC, but not in NLAC (4.0%); while there was no significant difference of IMP3 expression in LAC (88.2%) and NLAC (88.0%). The 487 bp DNA fragments of IMP3 was expected to be amplified in 6/9 of adenocarcinoma cases showed negative in ICC; and the 394 bp DNA fragments of CK5/6 was also expected to be amplified in 4/7 of RMs cases showed negative in ICC. Consistent with ICC results, there was significant difference of TTF1 expression in the LAC and NLAC compared with IMP3 expression. The combination with CK5/6, IMP3 and TTF1 immunostaining appears to be useful to improve the accuracy of cytological diagnoses between RMs, metastatic adenocarcinoma of lung and non-lung origin in pleural effusion. In addition, C-PCR may act as a useful supplemental approach for ICC, especially negative cases in ICC for differential cytological diagnosis.

IMP3 expression is associated with poor survival in cervical squamous cell carcinoma.

Insulin-like growth factor II messenger RNA (mRNA)-binding protein 3 (IMP3) is an oncofetal protein that promotes tumor progression and metastasis in a number of malignancies. However, the clinical significance of IMP3 expression in squamous cell carcinoma (SCC) of the uterine cervix is unclear. In this study, the correlation between IMP3 expression and cervical cancer progression and prognosis was assessed by immunohistochemistry. IMP3 expression was observed in a large number of tissue specimens from patients with normal cervical tissues, cervical intraepithelial neoplasia (CIN) I, CIN II, CIN III, and SCC. IMP3 protein and mRNA expression was also determined in the SiHa and HeLa human cervical cancer cell lines. IMP3 expression was observed in 0 (0%) of 62 CIN I, 0 (0%) of 38 CIN II, and 9 (8.7%) of 104 CIN III specimens. Of the 96 SCC cases, IMP3 expression was detected in 54 cases (56.3%). Significant difference in IMP3 expression existed between all of the groups tested (P < .001). IMP3 protein and mRNA expressions in SiHa and HeLa cell lines were dramatically increased, as compared with normal tissue (P < .001). IMP3 expression was significantly related to age (P < .001), International Federation of Gynecology and Obstetrics stage (P < .001), and lymph node metastasis (P = .001). IMP3 expression was also shown to be an independent prognostic factor in SCC. In conclusion, these findings suggest that IMP3 expression may be a prognostic indicator of SCC.

Combined detection of p16(INK4a) and IMP3 increase the concordance rate between cervical cytologic and histologic diagnosis.

Currently, there are discrepancies in the interpretation between cervical liquid-based cytology (LBC) and histologic diagnoses. The aim of our study was to evaluate the utility of p16(INK4a) (p16) and IMP3 staining of LBC specimens to increase the concordance rate. A total of 98 cell blocks with biopsy results, including 37 low-grade squamous intraepithelial lesions (LSIL), 36 high-grade squamous intraepithelial lesions (HSIL), and 25 squamous cell carcinomas (SCC), were selected for the immunocytochemical analysis of p16 and IMP3. The LBC diagnoses corresponded with histological diagnoses for 59.5% (22/37), 63.9% (23/36), and 88.0% (22/25) of LSIL, HSIL, and SCC lesions, respectively. We found a high frequency of p16 positivity in HSIL (72.2%) and SCC (100%), but not LSIL (29.7%). IMP3 was frequently expressed in SCC (84.0%), but rarely in LSIL (8.1%) and HSIL (25.0%). Cervical intraepithelial neoplasia 1 (CIN1) was negative for both p16 and IMP3, CIN2/3 tended to be positive for p16 and negative for IMP3, and SCC was positive for both p16 and IMP3. The combination of p16 and IMP3 immunostaining had a higher sensitivity and specificity for detecting CIN1 and CIN2/3 than cytology. For detecting SCC, p16/IMP3 had a higher sensitivity than cytology, but a lower specificity. IMP3 is a useful diagnostic immunomarker that can be used to identify SCC and the combination of p16/IMP3 expression was found to improve the discrepant results between cytologic and histologic diagnoses.

[Co-expression of CD99/MIC2 and ALK in anaplastic large-cell lymphoma tissues and its significance].

OBJECTIVE: To investigate co-expression of CD99/MIC2 and anaplastic lymphoma kinase (ALK) protein in anaplastic large-cell lymphoma (ALCL) tissues and Karpas 299 cells and its significance. METHODS: Clinical prognoses and ALK protein expressions of 25 cases of ALCL were reviewed retrospectively, the median duration of survival was analyzed for patients with ALK(+) ALCL and ALK(-) ALCL. Histological and immunohistochemical staining were applied to other 25 cases of ALCL and paraffin-embedded tissue from human anaplastic large-cell lymphoma Karpas 299 cells to detect the protein of CD99 and ALK. RESULTS: Of former 25 cases of ALCL, median duration of survival for ALK(+) patients was 59 months, whereas 20 months for ALK(-) patients. The prognosis of ALK(+) group was better than that of ALK(-) group, survival curves of these two groups showed statistically significant (P < 0.05). CD99 was positive in 18 cases (72.0%) while negative in 7 cases (28.0%) of the latter 25 ALCL, ALK was positive in 19 cases (76.0%) while negative in 6 cases (24.0%); Of 19 ALK(+) ALCL, 16 (84.2%) cases co-expressed CD99-ALK; and in 6 ALK(-) ALCL, 2(33.3%) were CD99-ALK double negative, the expression of CD99 protein strongly correlated with that of ALK protein (P < 0.05). ALK and CD99 protein expressed in Karpas 299 cells with diffuse distribution. CONCLUSIONS: CD99 highly expressed in ALCL, and showed high rate of co-expression with ALK. CD99 protein expression could be considered as a helpful diagnostic and prognostic factor of ALCL, especially for ALK(+) ALCL.