Effects of AZD8055 on autophagy and apoptosis in cholangiocarcinoma cells / 中国病理生理杂志

AIM:To explore the effects of mammalian target of rapamycin (mTOR) double inhibitor AZD8055 on autophagy and apoptosis of human cholangiocarcinoma cell line HuCCT1. METHODS:The effect of AZD8055 on the viability of HuCCT1 cells was detected by MTT assay. Autophagosome was detected by acridine orange (AO) staining. Af-ter treated with AZD8055, the expression levels of apoptosis-related proteins Bcl-2, Bax and cleaved caspase-3 and auto-phagy marker proteins beclin 1, LC3 and p62 were determined by Western blot. Apoptotic rate was analyzed by flow cyto-metry with Annexin V-FITC/PI double staining. RESULTS:AZD8055 significantly inhibited the viability of HuCCT1 cells (P<0.05). AO staining showed that AZD8055 significantly increased orange granules in the cytoplasm. After treated with AZD8055, compared with the control group, the protein level of beclin 1 and the ratio of LC3-Ⅱ/LC3-Ⅰ were enhanced, while p62 was attenuated (P<0.05). The protein expression level of pro-apoptotic regulator Bax was down-regulated and anti-apoptotic regulator Bcl-2 was increased. The protein level of cleaved caspase-3 was reduced (P<0.05). The results of flow cytometry showed that AZD8055 inhibited cell apoptosis. CONCLUSION:AZD8055 inhibits the viability of cholangiocarcinoma cells, and the mechanism is closely related with autophagy induced by AZD8055.

Baicalein inhibits proliferation and migration of gastric cancer MGC-803 cells / 中国病理生理杂志

AIM: To investigate the effects of baicalein(BAI)on the proliferation and migration of gastric cancer MGC-803 cells and the mechanisms.METHODS:After MGC-803 cells were treated with BAI at different concen-trations,the viability of the MGC-803 cells was tested by MTT assay.The cell colony formation ability were detected by plate colony formation assay.Wound-healing and Transwell cell migration assays were used to test the migration ability of the MGC-803 cells.The concentration of 12-hydroxyeicosatetraenoic acid(12-HETE)was measured by ELISA.The pro-tein levels of platelet type 12-lipoxygenase(p12-LOX),vascular endothelial growth factor(VEGF),p-ezrin and epithelial-mesenchymal transition(EMT)markers in MGC-803 cells were determined by Western blot.RESULTS:BAI significantly inhibited the proliferation,plate colony formation and migration abilities of the MGC-803 cells(P<0.05 or P<0.01), down-regulated the concentration of p12-LOX metabolite 12-HETE significantly(P<0.05 or P<0.01), decreased the protein levels of p12-LOX,VEGF,p-ezrin,vimentin and Snail(P<0.05 or P<0.01),and increased the protein expres-sion of E-cadherin(P<0.01).CONCLUSION:BAI suppresses the proliferation and migration abilities of gastric cancer MGC-803 cells effectively.These effects of BAI may be related to regulating the protein levels of p12-LOX,VEGF,p-ezrin and EMT-related proteins.

Relationship between hedgehog signaling pathway molecules and HPV16 infection in uterine cervical cancers / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the significance of sonic hedgehog (Shh), indian hedgehog (Ihh), smoothened (Smo) and patched (Ptch) expressions in uterine cervical lesions and their relationships with HPV type 16 infection.</p><p><b>METHODS</b>Totally 183 cases of cervical lesions, including 32 non-neoplastic cervix, 71 cervical intraepithelial neoplasia (28 CINI, 18 CINII, and 25 CINIII) and 80 squamous cell carcinomas (SCC) were selected from the Department of Pathology, Yanbian University Hospital, Yanbian Women Hospital, and Yanbian Tumor Hospital. Shh, Ihh, Ptch and Smo proteins expression were investigated by immunohistochemistry using tissue microarry platform, and the presence of HPV type 16 was detected by PCR method.</p><p><b>RESULTS</b>Immunohistochemical staining showed that the frequencies of Shh, Ihh, Ptch and Smo expression were rare in normal cervical epithelium, but were strongly expressed in cervical cancer and its precursor lesions (CINII/III) (P < 0.01, P < 0.01, P < 0.05, P < 0.05, respectively). In cervical cancer, the expression rate of Shh (95%) was higher than that of CIN (CINI to CINIII) (46.4%, 61.1%, 80.0%, respectively, P < 0.05). HPV16 was positive in 77.5% of SCC. In cervical cancer, the expression of Shh was related with HPV16 infection (P < 0.05), and the expression of Smo was correlated with lymph node metastasis (P < 0.05).</p><p><b>CONCLUSIONS</b>Shh, Ihh, Ptch, and Smo genes may play important roles in the development of cervical cancer. Detection of Hedgehog signaling pathway molecules seems helpful for the early diagnosis of cervical cancer and its precursor lesions, and are potentially therapeutic targets as well.</p>

Study of p16(INK4A) expression and DNA ploidy in HPV-negative cervical cancers and precursors / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathological significance of p16(INK4A) expression and DNA ploidy status in HPV-negative uterine cervical cancers and their precursors.</p><p><b>METHODS</b>HPV-negative cervical lesions, including 20 cases of cervicitis, 20 cases of cervical intraepithelial neoplasm (CIN), 3 cases of cervical glandular intraepithelial neoplasm (CGIN), 38 cases of invasive squamous cell carcinoma (SCCs) and 15 cases of invasive adenocarcinoma were selected and subject to screening for HPV infection by PCR method. The p16(INK4A) protein expression and DNA ploidy status were studied by immunohistochemistry and flow cytometry respectively.</p><p><b>RESULTS</b>Specific expression of p16(INK4A) was seen in both the nucleus and cytoplasm of the dysplastic and malignant cells of CIN, CGIN, cervical SCC and adenocarcinoma. In contrast, no expression was present in normal and inflammatory squamous or glandular epithelium. DNA aneuploidy was significantly more frequent in invasive SCCs and adenocarcinomas than in CIN (P < 0.01). Aneuploid was also more frequent in the lymph node positive group than lymph node negative group, although no statistic significance was found. Among the 8 cases of p16(INK4A) negative SCCs, two showed DNA aneuploidy.</p><p><b>CONCLUSIONS</b>Immunohistochemical detection for p16(INK4A) can be an early diagnostic marker for HPV-negative cervical SCC and adenocarcinoma. DNA ploidy analysis may further assist the diagnosis of cervical malignancies.</p>

Immunohistochemical Analysis of CD44s and CD44v6 in Endometriosis and Adenomyosis: Comparison with normal, hyperplastic, and malignant endometrium

The expression patterns of CD44s and CD44v6 were immunohistochemically compared with those of normal, hyperplastic and malignant endometrium. In normal endometria (n=37), endometrioses (n=46) and adenomyoses (n=20), the surface and glandular epithelial cells were negative for CD44s and CD44v6 in a proliferative pattern and positive in a secretory pattern, whereas the stroma was only positive for CD44s in both proliferative and secretory patterns. The endometrial hyperplasia (4 simple and 9 complex) had the identical patterns with normal proliferative phase of endometrium. Only one case showing complex hyperplasia with atypia was focally positive for CD44s and CD44v6 in glandular epithelia. CD44s and CD44v6 were positive in all endometrial adenocarcinomas (13), except one CD44s-negative case. In summary, the expressions of CD44s and CD44v6 in endometriosis and adenomyosis recapitulated those of normal cyclic endometrium. The expression patterns in endometrial hyperplasia were similar to those in normal proliferative endometrium, whereas the endometrial adenocarcinoma showed abnormal expressions for CD44s and CD44v6. Thus it was considered that the ectopic endometrium in endometriosis and adenomyosis was not aberrant as in endometrial carcinoma on the aspects of immunohistochemical expressions of CD44s and CD44v6.

Detection of Epstein-Barr virus in the inflammatory and neoplastic uterine cervical lesions / 대한세포병리학회지

The prevalence of Epstein-Barr virus(EBV) in the uterine cervix was investigated to define the possible etiologic role in cervical carcinogenesis. The viral genotyping and LMP-1 30bp deletion were also studied. The materials included 169 uterine cervical swabs(152 within normal limits, 12 atypical squamous cells of uncertain significance, 3 low grade intraepithelial lesions, and 2 high grade squamous intraepithelial lesion) and 104 uterine cervical tissues obtained from hysterectomy specimens(32 carcinoma in situ, 9 microinvasive squamous cell carcinomas, 37 invasive squamous cell carcinomas, 7 adenocarcinomas, 7 adenosquamous carcinomas, and 12 cervicitis). EBV detected by PCR for EBNA-1 was positive in 52(56.5%) of 92 invasive and noninvasive cervical carcinomas, and 80(48.8%) of 164 inflammatory or normal cervices. The viruses detected in carcinomas were all type A, and LMP-1 30bp deletion form was more frequent in premalignant and malignant cervical lesions than in nonneoplastic cervices. From the above results, it may be concluded that EBV is one of common viruses detected in uterine cervix of Korean women, and type A virus and LMP-1 30bp deletion form may have a role in cervical carcinogenesis.