Harel-Yoon syndrome caused by a novel variant in ATAD3A: A case report.

Objectives: To describe the clinical feature of a very recently identified phenotype associated with ATAD3A variation. Methods: A neonate with Harel-Yoon syndrome was identified. We describe the proband's clinical and radiological features. The affected newborn and her parents underwent whole-exome sequencing and PCR-Sanger sequencing. Results: Previously reported clinical manifestations were rare in the neonatal period, including unmanageable seizures necessitating the use of multiple drugs, congenital laryngeal stridor, hypotonia, challenges with feeding, corneal opacity, and subsequent demise due to respiratory failure. Molecular investigations have unveiled the presence of a newly identified heterozygous single-base substitution (c.1517A > C; p.Q506P) within the ATAD3A gene. Discussion: This study unveils a novel single-base substitution, thereby expanding the mutation spectrum associated with ATAD3A. Furthermore, the clinical characteristics exhibited during the neonatal phase are comprehensively described, potentially facilitating improved clinical recognition of ATAD3A-associated HAYOS.

Clinical features and imaging examination assessment of cervical lymph nodes for thyroid carcinoma.

BACKGROUNDS: The purpose of this study is to investigate the relationship between clinical characteristics and cervical lymph node metastasis (LNM) in patients with thyroid carcinoma, as well as estimate the preoperative diagnosis values of ultrasound (US) and contrast enhanced computed tomography (CECT) examinations on the neck for detection of cervical LNM in thyroid carcinoma. METHODS: A retrospective analysis of 3 026 patients with surgically proven thyroid carcinoma was conducted. Patients' clinical characteristics, including gender, age, tumor size, bilateral lesions, multifocality, adenomatous nodules, Hashimoto's thyroiditis (HT), and extrathyroidal extension, were collected to explore their association with cervical LNM in thyroid carcinoma. Preoperative assessments for central lymph node metastasis (CLNM) and lateral lymph node metastasis (LLNM) were conducted through US and CECT. The diagnostic value of US, CECT and US combined with CECT for detection of LNM located in various cervical compartments was estimated based on the pathological results. RESULTS: The risk of cervical LNM was higher in thyroid cancer patients who were male, age < 55 years old, tumor size > 10 mm, bilateral lesions, and extrathyroidal extension, while multifocality, adenomatous nodules and HT had no significant effect on LNM. US, CECT and US combined with CECT all had a higher sensitivity to LLNM (93.1%, 57.8%, 95.4%) than to CLNM (32.3%, 29.0%, 43.4%). US and CECT had a high specificity to both CLNM and LLNM (94.3-97.8%). CONCLUSION: Preoperative clinical characteristics and imaging examinations on patients with thyroid carcinoma are crucial to the evaluation of cervical lymph nodes and conducive to individualizing surgical treatments by clinicians. US combined with CECT are superior to single US or CECT alone in detection of CLNM and LLNM.

A traditional gynecological medicine inhibits ovarian cancer progression and eliminates cancer stem cells via the LRPPRC-OXPHOS axis.

BACKGROUND: Ovarian cancer (OC) is the most lethal malignant gynecological tumor type for which limited therapeutic targets and drugs are available. Enhanced mitochondrial oxidative phosphorylation (OXPHOS), which enables cell growth, migration, and cancer stem cell maintenance, is a critical driver of disease progression and a potential intervention target of OC. However, the current OXPHOS intervention strategy mainly suppresses the activity of the electron transport chain directly and cannot effectively distinguish normal tissues from cancer tissues, resulting in serious side effects and limited efficacy. METHODS: We screened natural product libraries to investigate potential anti-OC drugs that target OXPHOS. Additionally, LC-MS, qRT-PCR, western-blot, clonogenic assay, Immunohistochemistry, wound scratch assay, and xenograft model was applied to evaluate the anti-tumor mechanism of small molecules obtained by screening in OC. RESULTS: Gossypol acetic acid (GAA), a widely used gynecological medicine, was screened out from the drug library with the function of suppressing OXPHOS and OC progression by targeting the leucine-rich pentatricopeptide repeat containing (LRPPRC) protein. Mechanically, LRPPRC promotes the synthesis of OXPHOS subunits by binding to RNAs encoded by mitochondrial DNA. GAA binds to LRPPRC directly and induces LRPPRC rapid degradation in a ubiquitin-independent manner. LRPPRC was overexpressed in OC, which is highly correlated with the poor outcomes of OC and could promote the malignant phenotype of OC cells in vitro and in vivo. GAA management inhibits cell growth, clonal formation, and cancer stem cell maintenance in vitro, and suppresses subcutaneous graft tumor growth in vivo. CONCLUSIONS: Our study identified a therapeutic target and provided a corresponding inhibitor for OXPHOS-based OC therapy. GAA inhibits OC progression by suppressing OXPHOS complex synthesis via targeting LRPPRC protein, supporting its potential utility as a natural therapeutic agent for ovarian cancer.

A Nomogram for Enhancing the Diagnostic Effectiveness of Solid Breast BI-RADS 3-5 Masses to Determine Malignancy Based on Imaging Aspects of Conventional Ultrasonography and Contrast-Enhanced Ultrasound.

BACKGROUND: To establish and validate a nomogram model, which can incorporate clinical data, and imaging features of ultrasound (US) and contrast-enhanced ultrasound (CEUS), for improving the diagnostic efficiency of solid breast lesions. PATIENTS AND METHODS: A total of 493 patients with solid breast lesions were randomly divided into training (n = 345) and validation (n = 148) cohorts with a ratio of 7:3 and, clinical data and image features of US and CEUS were reviewed and retrospectively analyzed. The breast lesions in both the training and validation cohorts were analyzed using the BI-RADS and nomogram models. RESULTS: Five variables, including the shape and calcification features of conventional US, enhancement type and size after enhancement features of CEUS, and BI-RADS, were selected to construct the nomogram model. As compared to the BI-RADS model, the nomogram model demonstrated satisfactory discriminative function (area under the receiver operating characteristic [ROC] curves [AUC], 0.940; 95% confidence interval [CI], 0.909 to 0.971; sensitivity, 0.905; and specificity, 0.902 in the training cohort and AUC, 0.968; 95% CI, 0.941 to 0.995; sensitivity, 0.971; and specificity, 0.867 in the validation cohort). In addition, the nomogram model showed good consistency and clinical potential according to the calibration curve and DCA. CONCLUSION: The nomogram model could identify benign from malignant breast lesions with good performance.

Combined Conventional Ultrasound and Contrast-Enhanced Computed Tomography for Cervical Lymph Node Metastasis Prediction in Papillary Thyroid Carcinoma.

OBJECTIVES: This study aimed to evaluate conventional ultrasound (US) combined with contrast-enhanced computed tomography (CT) of the neck to predict central lymph node metastasis (CLNM) in clinical lymph-negative patients with papillary thyroid carcinoma (PTC), establish a simple preoperative risk-scoring model, and validate its effectiveness in a two-center dataset. METHODS: A total of 423 patients with PTC preoperatively evaluated by US and contrast-enhanced CT were included in the modeling group, and 102 patients from two hospitals were enrolled in the validation group. Independent predictive factors were determined using multivariate logistic regression analysis. Diagnostic performance was evaluated using receiver operating characteristic curve analysis. RESULTS: The independent predictive factors for CLNM were age ≤45 years (odds ratio [OR] = 3.950), nodule presence in the non-upper pole (OR = 2.385), nodule size >12.5 mm (OR = 2.130), Thyroid Imaging Reporting and Data System score ≥9 (OR = 2.857), normalized enhancement CT value ≥0.75 (OR = 3.132), central enhancement (OR = 0.222), and capsular invasion (OR = 3.478). The area under the curve (AUC) of the model was 0.790 (95% confidence interval [CI]: 0.747-0.834), and the sensitivity and specificity were 70.4% and 73.9%, respectively. The AUC in the validation group was 0.827 (95% CI: 0.747-0.907), and the sensitivity and specificity were 88.9% and 63.2%, respectively. CONCLUSIONS: We found conventional US combined with contrast-enhanced CT of the neck to be useful in predicting CLNM preoperatively and established a simple risk-scoring model that might help surgeons with appropriate surgical plans and prognostic evaluation.

An artificial intelligence ultrasound system's ability to distinguish benign from malignant follicular-patterned lesions.

Objectives: To evaluate the application value of a generally trained artificial intelligence (AI) automatic diagnosis system in the malignancy diagnosis of follicular-patterned thyroid lesions (FPTL), including follicular thyroid carcinoma (FTC), adenomatoid hyperplasia nodule (AHN) and follicular thyroid adenoma (FTA) and compare the diagnostic performance with radiologists of different experience levels. Methods: We retrospectively reviewed 607 patients with 699 thyroid nodules that included 168 malignant nodules by using postoperative pathology as the gold standard, and compared the diagnostic performances of three radiologists (one junior, two senior) and that of AI automatic diagnosis system in malignancy diagnosis of FPTL in terms of sensitivity, specificity and accuracy, respectively. Pairwise t-test was used to evaluate the statistically significant difference. Results: The accuracy of the AI system in malignancy diagnosis was 0.71, which was higher than the best radiologist in this study by a margin of 0.09 with a p-value of 2.08×10-5. Two radiologists had higher sensitivity (0.84 and 0.78) than that of the AI system (0.69) at the cost of having much lower specificity (0.35, 0.57 versus 0.71). One senior radiologist showed balanced sensitivity and specificity (0.62 and 0.54) but both were lower than that of the AI system. Conclusions: The generally trained AI automatic diagnosis system can potentially assist radiologists for distinguishing FTC from other FPTL cases that share poorly distinguishable ultrasonographical features.

The diagnostic value of a nomogram based on multimodal ultrasonography for thyroid-nodule differentiation: A multicenter study.

Objective: To establish and verify a nomogram based on multimodal ultrasonography (US) for the assessment of the malignancy risk of thyroid nodules and to explore its value in distinguishing benign from malignant thyroid nodules. Methods: From September 2020 to December 2021, the data of 447 individuals with thyroid nodules were retrieved from the multicenter database of medical images of the National Health Commission's Capacity Building and Continuing Education Center, which includes data from more than 20 hospitals. All patients underwent contrast-enhanced US (CEUS) and elastography before surgery or fine needle aspiration. The training set consisted of three hundred datasets from the multicenter database (excluding Zhejiang Cancer Hospital), and the external validation set consisted of 147 datasets from Zhejiang Cancer Hospital. As per the pathological results, the training set was separated into benign and malignant groups. The characteristics of the lesions in the two groups were analyzed and compared using conventional US, CEUS, and elastography score. Using multivariate logistic regression to screen independent predictive risk indicators, then a nomogram for risk assessment of malignant thyroid nodules was created. The diagnostic performance of the nomogram was assessed utilizing calibration curves and receiver operating characteristic (ROC) from the training and validation cohorts. The nomogram and The American College of Radiology Thyroid Imaging, Reporting and Data System were assessed clinically using decision curve analysis (DCA). Results: Multivariate regression showed that irregular shape, elastography score (≥ 3), lack of ring enhancement, and unclear margin after enhancement were independent predictors of malignancy. During the training (area under the ROC [AUC]: 0.936; 95% confidence interval [CI]: 0.902-0.961) and validation (AUC: 0.902; 95% CI: 0.842-0.945) sets, the multimodal US nomogram with these four variables demonstrated good calibration and discrimination. The DCA results confirmed the good clinical applicability of the multimodal US nomogram for predicting thyroid cancer. Conclusions: As a preoperative prediction tool, our multimodal US-based nomogram showed good ability to distinguish benign from malignant thyroid nodules.

Differential diagnosis of non-diffuse primary thyroid lymphoma and papillary thyroid carcinoma by ultrasound combined with computed tomography.

BACKGROUND: Primary thyroid lymphoma (PTL) and papillary thyroid carcinoma (PTC) are both thyroid malignancies, but their therapeutic methods and prognosis are different. This study aims to explore their sonographic and computed tomography(CT)features, and to improve the early diagnosis rate. METHODS: The clinical and imaging data of 50 patients with non-diffuse PTL and 100 patients with PTC confirmed by pathology were retrospectively analysed. RESULTS: Of the 150 patients, from the perspective of clinical data, between non-diffuse PTL and PTC patients existed significant difference in age, maximum diameter of nodule, asymmetric enlargement and Hashimoto's thyroiditis (P < 0.001), but not in gender ratio, echo texture, cystic change and anteroposterior-to-transverse ratio (P > 0.05). With respect to sonographic feature, non-diffuse PTL patients had a higher proportion than PTC patients in markedly hypoechoic, internal linear echogenic strands, posterior echo enhancement, rich vascularity, lack of calcification and homogeneous enhancement, with statistically significant difference (P < 0.05), while PTC patients had a higher proportion than non-diffuse PTL patients in irregular border, circumscribed margin, capsular invasion and significant enhancement, with statistically significant difference (P < 0.001). With respect to CT feature, non-diffuse PTL patients were significantly different from PTC patients in the non-contrast CT value mean, venous phase CT value mean, enhanced intensity and homogeneity of nodules (P < 0.05). Multivariate logistic regression analysis showed that age (OR = 1.226, 95%CI:1.056 ~ 1.423, P = 0.007), posterior echo enhancement (OR = 51.152, 95%CI: 2.934 ~ 891.738, P = 0.007), lack of calcification (OR = 0.013, 95%CI: 0.000 ~ 0.400, P = 0.013) and homogeneous enhancement (OR = 0.020, 95%CI: 0.001 ~ 0.507, P = 0.018) were independent risk factors. CONCLUSIONS: Sonographic and CT features of the presence of posterior echo enhancement, lack of calcification and homogeneous enhancement were valuable to distinguishing non-diffuse PTL from PTC.

Multimodal Sonographic Appearance and Survival Outcomes of 69 Cases of Primary Thyroid Lymphoma Over 10 Years.

OBJECTIVES: To investigate ultrasound appearance and the survival outcomes for patients with primary thyroid lymphoma (PTL). METHODS: Ultrasonic images and clinical characteristics from pathologically confirmed 69 PTL patients (2008-2019) were retrospectively analyzed. The clinical characteristics, ultrasonic characters, and prognostic factors were analyzed. Survival curves were plotted using the Kaplan-Meier method. Univariate and multivariate analyses were performed. RESULTS: Of the 69 study patients, 23 were indolent PTL and 46 were aggressive PTL. Age (>70 years old) and elevated lactate dehydrogenase levels were statistically different clinical features between aggressive and indolent PTL. From ultrasonic images, 34 cases were nodular, 11 diffuse, and 24 mixed pattern. Mixed types displayed high invasiveness (45.7%) while diffuse types displayed higher inertness (39.1%), with statistically significant differences (P = .000). Invaded thyroid capsule and increased chaotic vascularity also showed significant differences between aggressive and indolent PTL. We also observed statistical difference in overall survival rates between aggressive and indolent PTL (P = .032). Single factor K-M analyses showed that age >70 years, aggressive pathology, and Ki67 >30% were positively correlated with the risk of poor PTL survival (P < .05). CONCLUSIONS: Multimodal ultrasound provides accurate ultrasonographic information and facilitates PTL invasiveness diagnostics for improved clinical treatment. In addition, PTL patients aged >70 years, with aggressive pathology, and Ki67 >30% were more likely to have a poor survival outcome.

Value of deep learning models based on ultrasonic dynamic videos for distinguishing thyroid nodules.

Objective: This study was designed to distinguish benign and malignant thyroid nodules by using deep learning(DL) models based on ultrasound dynamic videos. Methods: Ultrasound dynamic videos of 1018 thyroid nodules were retrospectively collected from 657 patients in Zhejiang Cancer Hospital from January 2020 to December 2020 for the tests with 5 DL models. Results: In the internal test set, the area under the receiver operating characteristic curve (AUROC) was 0.929(95% CI: 0.888,0.970) for the best-performing model LSTM Two radiologists interpreted the dynamic video with AUROC values of 0.760 (95% CI: 0.653, 0.867) and 0.815 (95% CI: 0.778, 0.853). In the external test set, the best-performing DL model had AUROC values of 0.896(95% CI: 0.847,0.945), and two ultrasound radiologist had AUROC values of 0.754 (95% CI: 0.649,0.850) and 0.833 (95% CI: 0.797,0.869). Conclusion: This study demonstrates that the DL model based on ultrasound dynamic videos performs better than the ultrasound radiologists in distinguishing thyroid nodules.

Combination of transcatheter arterial chemoembolization and portal vein embolization for patients with hepatocellular carcinoma: a review.

BACKGROUND: Transcatheter arterial chemoembolization has been widely used in patients with hepatocellular carcinoma. However, double blood supply and the existence of portal vein tumor thrombus influence the efficacy of transcatheter arterial chemoembolization. MAIN BODY: Theoretically, portal vein embolization combined with transcatheter arterial chemoembolization may bring a breakthrough in the therapeutic effect of hepatocellular carcinoma. The feasibility, efficacy, long-term survival benefits, and side effects of the combined treatment have been explored in previous studies. Chemotherapeutic agents may also be added in the portal vein embolization procedure to further improve the treatment response. CONCLUSION: In this study, we review the existing data and studies on the combined treatment in patients with hepatocellular carcinoma and provide an overall view of the strategy.

In Situ Bismuth Nanosheet Assembly for Highly Selective Electrocatalytic CO2 Reduction to Formate.

The reduction of carbon dioxide (CO2 ) into value-added fuels using an electrochemical method has been regarded as a compelling sustainable energy conversion technology. However, high-performance electrocatalysts for CO2 reduction reaction (CO2 RR) with high formate selectivity and good stability need to be improved. Earth-abundant Bi has been demonstrated to be active for CO2 RR to formate. Herein, we fabricated an extremely active and selective bismuth nanosheet (Bi-NSs) assembly via an in situ electrochemical transformation of (BiO)2 CO3 nanostructures. The as-prepared material exhibits high activity and selectivity for CO2 RR to formate, with nearly 94% faradaic efficiency at -1.03 V (versus reversible hydrogen electrode (vs. RHE)) and stable selectivity (>90%) in a large potential window ranging from -0.83 to -1.18 V (vs. RHE) and excellent durability during 12 h continuous electrolysis. In addition, the Bi-NSs based CO2 RR/methanol oxidation reaction (CO2 RR/MOR) electrolytic system for overall CO2 splitting was constructed, evidencing the feasibility of its practical implementation.

Hydrangea-like Superstructured Micro/Nanoreactor of Topotactically Converted Ultrathin Bismuth Nanosheets for Highly Active CO2 Electroreduction to Formate.

An electrocatalytic carbon dioxide reduction reaction (CO2RR) is an appealing route to obtain the value-added feedstocks and alleviate the energy crisis. However, how to achieve high-performance electrocatalysts for CO2 reduction to formate is challenging owing to the poor intrinsic activity, insufficient conductivity, and low surface density of active sites. Herein, we fabricated an extremely active and selective hydrangea-like superstructured micro/nanoreactor of ultrathin bismuth nanosheets through an in situ electrochemical topotactic transformation of hierarchical bismuth oxide formate (BiOCOOH). The resulted bismuth nanosheet superstructure is in the form of three-dimensional intercrossed networks of ultrathin nanosheets, forming an ordered open porous structure through self-assembly, which can be used as a micro/nanoreactor to enable a large electrochemically active surface area as well as high atomic utilization. Such a distinctive nanostructure endows the material with high electrocatalytic performances for CO2 reduction to formate with near-unity Faradaic selectivity (>95%) in a wide potential window from -0.78 to -1.18 V. Furthermore, this micro/nanoreactor can give the high current densities over 300 mA cm-2 at low applied potentials without compromising selectivity in a flow cell reactor. Density functional theory (DFT) and in situ attenuated total reflection-infrared spectroscopy (in situ ATR-IR) were further conducted to interpret the CO2RR mechanisms.

Using ultrasound-targeted microbubble destruction to enhance radiotherapy of glioblastoma.

OBJECTIVE: To investigate the efficacy and mechanism of ultrasound-targeted microbubble destruction (UTMD) combined with radiotherapy (XRT) on glioblastoma. METHODS: The enhanced radiosensitization by UTMD was assessed through colony formation and cell apoptosis in Human glioblastoma cells (U87MG). Subcutaneous transplantation tumors in 24 nude mice implanted with U87MG cells were randomly assigned to 4 different treatment groups (Control, UTMD, XRT, and UTMD + XRT) based on tumor sizes (100-300 mm3). Tumor growth was observed for 10 days after treatment, and then histopathology stains (HE, CD34, and γH2AX) were applied to the tumor samples. A TUNEL staining experiment was applied to detect the apoptosis rate of mice tumor samples. Meanwhile, tissue proteins were extracted from animal specimens, and the expressions of dsDNA break repair-related proteins from animal specimens were examined by the western blot. RESULTS: When the radiotherapy dose was 4 Gy, the colony formation rate of U87MG cells in the UTMD + XRT group was 32 ± 8%, lower than the XRT group (54 ± 14%, p < 0.01). The early apoptotic rate of the UTMD + XRT group was 21.1 ± 3% at 48 h, higher than that of the XRT group (15.2 ± 4%). The tumor growth curve indicated that the tumor growth was inhibited in the UTMD + XRT group compared with other groups during 10 days of observation. In TUNEL experiment, the apoptotic cells of the UTMD + XRT group were higher than that of the XRT group (p < 0.05). The UTMD + XRT group had the lowest MVD value, but was not significantly different from other groups (p > 0.05). In addition, γH2AX increased due to the addition of UTMD to radiotherapy compared to XRT in immunohistochemistry (p < 0.05). CONCLUSIONS: Our study clearly demonstrated the enhanced destructive effect of UTMD combined with 4 Gy radiotherapy on glioblastoma. This could be partly achieved by the increased ability of DNA damage of tumor cells.

Biplane transrectal ultrasonography plus ultrasonic elastosonography and contrast-enhanced ultrasonography in T staging of rectal cancer.

BACKGROUND: The aim of this study is to assess biplane transrectal ultrasonography (TRUS) plus ultrasonic elastosonography (UE) and contrast-enhanced ultrasonography (CEUS) in T staging of rectal cancer. METHODS: Between March 2016 and January 2019, 66 rectal cancer patients who completed biplane TRUS plus UE and CEUS for preoperative workup and were treated by primary total mesorectal excision (TME) were retrospectively analyzed. RESULTS: The accuracy of TRUS plus UE and CEUS in all T staging of rectal cancer was 69.7%. The highest accuracy was achieved in the T3 stage (87.5%), while it was 71.4 and 50.0% in the T1 and T2 stage, respectively. The mean sizes of uT1-T2 lesions and uT3-T4 lesions were 30.0 ± 10.6 mm (range, 10.0-55.0) and 40.2 ± 11.2 mm (range, 14.0-57.0), respectively (p < 0.001). According to the receiver operating characteristic (ROC) curve to predict pT stages (pT1,2 vs. pT3), the optimal cut-off value of lesions in greatest dimension was 28.5 mm by TRUS with areas under the curve (AUC) of 0.769, and the optimal cut-off values of peak systolic velocity (PSV) and resistive index (RI) were 18.8 cm/sec and 0.645, respectively. The AUCs of PSV and RI were 0.588 and 0.555, respectively. CONCLUSIONS: Diagnostic accuracy of TRUS plus UE and CEUS in T staging of rectal cancer does not reach the excellent published study results, especially for patients with early rectal cancer. Tumor sizes, PSV and RI are useful additions for TRUS in T staging of rectal cancer.

Inhibition of cervical cancer cell growth in vitro and in vivo by lentiviral-vector mediated shRNA targeting the common promoter of HPV16 E6 and E7 oncogenes.

Deregulated expression of high-risk human papillomavirus oncogenes (E6 and E7) is a pivotal event for pathogenesis and progression in cervical cancer. Both viral oncogenes are therefore regarded as ideal therapeutic targets. Small interfering RNAs (siRNA) or double-stranded RNAs can knock down target genes effectively through siRNA-induced transcriptional gene silencing (TGS). Here, we established lentiviral-vector mediated shRNA (LV-shRNA) targeting common promoter of HPV16 E6/E7 and targeting E6 transcript, transduced the lentiviral construct into cervical HPV16-positive cell lines Siha and Caski, then selected and established stably transduced monoclonal cell lines. The results showed that LV-shRNA targeting promoter, as well as targeting E6 transcript, effectively knocked down E6 and E7 expression, resulted in accumulation of p53 and pRB protein and decrease of MCM7 and p16 protein, and consequently remarkably reduced the abilities of proliferation and invasiveness of cervical cancers cells in vitro. Then we inoculated subcutaneously those monoclonal cells into nude mice to establish the transplanted tumor animal models, and found dramatically inhibited tumorigenesis and growth, as well as prolonged survival time of mice incubated by cells with LV-shRNA targeting promoter and E6 transcript. Our results may provide evidence for application of LV-shRNA targeting HR-HPV key oncogenes, as a new treatment strategy, in cervical and other HPV-associated cancer therapy.

Transcriptional gene silencing of HPV16 E6/E7 induces growth inhibition via apoptosis in vitro and in vivo.

OBJECTIVE: Transcriptional silencing of HPV oncogenes using short interfering RNA (siRNA) blocks E6/E7 expression. Our objective was to estimate the effective value of E6/E7 specific siRNA-induced transcriptional gene silencing as a potential therapeutic strategy for cervical cancer. METHODS: In vitro studies were performed by employing two categories of siRNA targeting promoter of E6/E7 gene and E7 transcript, respectively, and inhibitory effect of both siRNAs was further observed in vitro and on xenograft in BALB/c mice that were inoculated with siRNA transfected SiHa cells and parental SiHa cells followed by siRNA intratumoral injection in vivo. Tumor volume and growth curves were assessed. Furthermore, cellular proliferation and apoptosis of inoculated tumors were determined by immunohistochemistry staining and TUNEL assay. RESULTS: The two most active siRNA sequences specifically knockdown E6/E7 expressions at mRNA level in HPV16 positive Siha cells, increased p53 and decreased p16 expressions at protein level, inhibited cell proliferation, and induced cell apoptosis in vitro. Furthermore, both siRNAs effectively inhibited tumor formation and growth no matter in mice with siRNA transfected cells in vitro or with siRNA intratumoral injection in vivo. TUNEL staining and FCM assay consistently showed that tumor retardation was through induction of cellular apoptosis. CONCLUSION: RNAi targeting the promoter of HPV16 E6/E7 acts effectively in vitro and in vivo, especially through intratumoral delivery, and may be a candidate therapeutic strategy for cervical cancer.

miR-375 is down-regulated in squamous cervical cancer and inhibits cell migration and invasion via targeting transcription factor SP1.

Pelvic lymph node metastases are regarded as the most important risk factor and a predictor of poor prognosis for patients with cervical cancer. Exploration of metastasis-related molecules is helpful toward improving the prognosis in cervical cancer. To identify the role of miR-375 in metastasis and progression of cervical cancer, we examined the expression of miR-375 in 170 cervical cancer tissues and 68 normal cervical tissues, using stem-loop quantitative PCR, and found that the expression of miR-375 in cervical cancer tissues was significantly decreased by 4.45-fold, compared with 68 normal tissues. A significant correlation existed between miR-375 expression and clinicopathologic parameters, including lymph node metastasis of cervical cancer. Overexpressed miR-375 suppressed cell proliferation, blocked G1-to-S cell-cycle transition, and inhibited cell migration and invasion in human cervical SiHa and CaSki cells. SP1, a potential target gene of miR-375, was inversely correlated with miR-375 expression in cervical cancer tissues. Moreover, SP1 was negatively regulated by miR-375, and knockdown of SP1 by siRNA inhibited cell malignant behaviors. Thus, our findings suggest that down-regulated miR-375 promotes cell malignant behaviors via the target gene SP1 and may consequently contribute to the progression of cervical cancer.