2.Role and action mechanism of aquaporin 3 in alleviating photoaging of skin fibroblasts by regulating hnRNPQ/p53
Huaxiong ZHANG ; Sha YAN ; Lin HE ; Lin LI ; Zhaohui CHEN ; Ji LI
Chinese Journal of Dermatology 2021;54(4):325-334
Objective:To investigate the role and action mechanism of aquaporin 3 (AQP3) in skin photoaging.Methods:Normal human skin fibroblasts (NHDF) were divided into several groups: NHDF group receiving normal culture without transfection, AQP3 cDNA group transfected with AQP3 cDNA, AQP3 siRNA group transfected with AQP3 siRNA, heterogeneous nuclear ribonucleoprotein Q (hnRNPQ) cDNA group transfected with hnRNPQ cDNA, hnRNPQ siRNA group transfected with hnRNPQ siRNA, AQP3-hnRNPQ cDNA group transfected with AQP3 and hnRNPQ cDNAs, AQP3-hnRNPQ siRNA group transfected with AQP3 and hRNPQ siRNAs, cDNA empty vector group transfected with a cDNA empty vector, and siRNA empty vector group transfected with a siRNA empty vector. Transfected or untransfected NHDFs were irradiated with ultraviolet A (UVA) at a dose of 10 J·cm -2·d -1 for 3 consecutive days to establish a model of cellular senescence, and NHDF receiving no UVA irradiation served as a control. A cell counting method was used to evaluate the cellular proliferative activity, a senescence-related β-galactosidase staining kit to evaluate the senescence level of NHDFs in each experimental group, and luciferase reporter gene technology to assess the transcriptional regulation activity of p53. Western blot analysis was performed to determine the expression of AQP3, hnRNPQ and senescence-related proteins p53 and p21 in NHDFs. Two-independent-sample t test was used for comparisons between two groups, and one-way analysis of variance for comparisons among multiple groups. Results:After 3-day consecutive irradiation with UVA, the expression of p53 and p21 in NHDFs and the percentage of β-galactosidase-positive cells significantly increased compared with the unirradiated control group (all P < 0.05) , but the expression of AQP3 and cellular proliferative activity on days 5, 6 and 7 significantly decreased in the UVA group compared with the unirradiated control group (all P < 0.05) . After 3-day consecutive irradiation with UVA, aggravated senescence-related phenotypes of UVA-induced NHDFs were observed in the AQP3 siRNA group compared with the siRNA empty vector group, and there were significant differences in the expression of p53, p21 and hnRNPQ, percentage of β-galactosidase-positive cells, p53 transcriptional regulation activity and cellular proliferative activity between the 2 groups (all P < 0.05) . Further silencing of the hnRNPQ gene could reverse the above effects. Compared with the siRNA empty vector group, the senescence-related phenotypes of UVA-induced NHDFs were attenuated in the hnRNPQ siRNA group, and significant differences were observed between the 2 groups in terms of the expression of p53, p21 and hnRNPQ, percentage of β-galactosidase-positive cells, p53 transcriptional regulation activity and cellular proliferative activity (all P < 0.05) . After 3-day consecutive irradiation with UVA, the senescence-related phenotypes of UVA-induced NHDFs were significantly attenuated in the AQP3 cDNA group compared with the cDNA empty vector group (all P < 0.05) , manifesting as significantly decreased expression of p53 (0.25 ± 0.06 vs. 0.56 ± 0.08) , p21 (0.23 ± 0.06 vs. 0.70 ± 0.07) and hnRNPQ (0.82 ± 0.09 vs. 0.92 ± 0.03) , percentage of β-galactosidase-positive cells (31.23% ± 6.54% vs. 81.53% ± 7.62%) and p53 transcriptional regulation activity (2.52 ± 0.36 vs. 7.16 ± 0.25) , but increased cellular proliferative activity ([2.93 ± 0.33]× 10 6/ml vs.[2.15 ± 0.23]× 10 6/ml) , and further overexpression of hnRNPQ could reverse the above effects. After 3-day consecutive irradiation with UVA, the expression of p53, p21, percentage of β-galactosidase-positive cells, p53 transcriptional regulation activity and cellular proliferative activity in the hnRNPQ cDNA group were 1.41 ± 0.09, 1.42 ± 0.06, 91.06% ± 4.24%, 12.35 ± 0.88 and (1.23 ± 0.41) × 10 6/ml respectively, and the senescence-related phenotypes of UVA-induced NHDFs were significantly aggravated in the hnRNPQ cDNA group compared with the cDNA empty vector group (all P < 0.05) . Conclusion:AQP3 may alleviate the UVA-induced senescence of NHDFs by regulating hnRNPQ and downregulating p53 expression.
3.Expression and significance of Twist and E-cadherin and Vimentin in cervical squamous epithelial cancerization
Jianbing HU ; Jing LIU ; Wen XIE ; Yong YANG ; Huaxiong GUO ; Jun LUO ; Weiwei GUO ; Ping ZHANG ; Peng ZHANG
Journal of Chinese Physician 2012;14(4):473-476
ObjectiveTo explore the expression rule and clinic significance of Twist and E-cadherin and Vimentin in cervical squamous epithelial cancerization.MethodsChronic cervicitis were used as control group,the expression of Twist (using hybridization in situ),E-cadherin and Vimentin( SABC immunohistochemical stain) in tissues of cervical intraepithelial neoplasia (CIN) and cervical squamous carcinoma were detected,and the correlations were analyzed.ResultsThe positive rate of Twist in group of chronic cervicitis,CIN Ⅰ,CINⅡ,CIN Ⅲ and cervical squamous carcinoma was 10.0%,28.1%,29.6%,42.9% and 64.3%,respectively.The expression rate of Twist in cervical squamous carcinoma group was higher than other groups ( P < 0.05 ).The positive rates of E-cadherin and Vimentin were 80.0%,68.8%,55.6%,51.4%,39.3% and 0,6.3%,7.4%,31.4%,32.1%,respectively.The difference of E-cadherin and Vimentin expression between the cervical squamous carcinoma ( or CIN Ⅲ group) and the chronic cervicitis group was obvious ( P <0.05,respectively).The expression of Twist was negatively correlated with E-cadherin( r =-0.37,P <0.01 ),and it was positively correlated with Vimentin in all cases of CIN and cervical squamous carcinoma( r =0.23,P <0.05).ConclusionsThere is a close relationship between abnormal expression of Twist and cervical squamous epithelial cancerization.Twist may participate in the genesis of cervical squamous carcinoma through mediating the expression of E-cadherin and Vimentin in cervix tissue.
4.Features of CT imaging and clinicopathological characteristics in multilocular cystic re-nal neoplasm of low malignant potential and cystic nephroma
Ling ZHANG ; Huaxiong GUO ; Ping GONG ; Xiuheng LIU ; Hao SHEN
Chinese Journal of Clinical Oncology 2019;46(13):669-672
Objective: To explore the clinicopathological features and the diagnostic value of the CT scan in multilocular cystic renal neoplasm of low malignant potential (MCRNLMP) and cystic nephroma (CN). Methods: The clinical data of 12 patients with CN and 20 patients with MCRNLMP, confirmed by pathology at the Renmin Hospital of Wuhan University and Jingzhou Central Hospital from Janu-ary 2000 to March 2019, were retrospectively analyzed. The receiver operating characteristic (ROC) curves were used to analyze the feature of contrast-enhanced CT images of the tumors, and the immunophenotypes of the tumors were observed by immunohisto-chemistry. Results: There were statistically significant differences between MCRNLMP and CN in terms of thickness of the cyst wall and partition, number of soft-tissue enhancing masses, peak intensity of enhancement, and the Bosniak classification (P<0.05). Based on ROC curve analyses, when the thickness of the capsule wall and partition was greater than 2.25mm, the number of enhanced high-density lesions was greater than 1, and the peaking intensity of fortification was above the moderate level in the diagnosis of CRNLMP. The areas under the curve of the three indexes were 0.879, 0.800 and 0.838, which can be used as the best diagnostic criterion for MCRNLMP. Immunophenotyping revealed that MCRNLMP characteristically expressed the renal cell carcinoma (RCC) marker, and CN characteristically expressed the estrogen receptor(ER) and progesterone receptor(PR). Conclusions: The cyst wall and septal thickness, number of soft-tissue enhancing masses, and peak intensity of enhancement show a higher diagnostic value in differentiating MCRN-LMP and CN. The precise diagnosis relies on the pathological and immunohistochemical examination.