Detection of genes responsible for cetuximab sensitization in colorectal cancer cells using CRISPR-Cas9.

Colorectal cancer (CRC) is a common malignant tumor in digestive tract with highly invasive and metastatic capacity. Drug sensitivity remains a significant obstacle to successful chemotherapy in CRC patients. The present study aimed to explore genes related to cetuximab (CTX) sensitivity in CRC by clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9. Celigo image cytometer was used to detect suitable cells and optimal dosage of CTX. Inhibition rate of CTX on Caco-2 cells was evaluated by cell counting kit-8 (CCK-8) method before and after transfection. 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide (MTT) was performed to explore suitable concentration of puromycin and multiplicity of infection (MOI). CRISPR-Cas9, sequencing data quality analysis and cell viability test were used for the selection of genes related to CTX sensitivity in CRC cells. Finally, the selected genes associated with CTX sensitivity in CRC cells were further validated by colony formation and CCK-8 assays. In the present study, Caco-2 cells had a better prolificacy, and CTX 100 µg/ml exhibited a good inhibition trend on the 7th and 14th days of infection. MTT assay indicated that the minimum lethal concentration of puromycin was 2.5 µg/ml. Forty-six candidate genes were preliminarily screened via sequencing data quality analysis. Subsequently, we found that knockout of any of the four genes (MMP15, MRPL48, CALN1 and HADHB) could enhance CTX sensitivity in Caco-2 cells, which was further confirmed by colony formation assay. In summary, MMP15, MRPL48, CALN1 and HADHB genes are related to the mediation of CTX sensitivity in CRC.

Utility of Red Cell Distribution Width as a Prognostic Factor in Young Breast Cancer Patients.

The prognosis of breast cancer occurs in young women is usually poor. Red cell distribution width (RDW), 1 of many routinely examined parameters, has recently been proposed as a prognostic marker in solid tumors. The aim of our study was to assess the predictive value of RDW for survival in young women with breast cancer.We reviewed 203 consecutive young female patients (under 40) with invasive breast cancer diagnosed at the First Affiliated Hospital of Wenzhou Medical University between January 2008 and December 2012. Preoperational RDW, clinicopathological information, and prognostic data were collected. RDW levels were divided into 2 groups: 161 patients with low RDW (≤13.75%) and 42 patients with high RDW (>13.75%). Clinicopathological differences between the 2 groups were calculated by chi-squared test and Wilcoxon rank-sum test. Kaplan-Meier survival analysis and Cox proportional hazard regression analyses were used to examine the effect of RDW on survival.We found that high RDW was significantly associated with larger tumor size (P = 0.002), positive lymph node metastases (P = 0.011), and advanced stages (P = 0.004). Patients with high RDW showed significantly lower disease-free survival (DFS; P < 0.001) and lower overall survival (OS) rate (P < 0.001) than patients with low RDW. Moreover, the Cox regression multivariate analysis revealed that high pretreatment DRW was independently correlated with poor DFS and OS, with hazard ratio 4.819 (95% confidence interval [CI] 2.291-10.138, P < 0.001) and 5.887 (95% CI 1.666-20.802, P = 0.006), respectively.In conclusion, our study demonstrated that pretreatment RDW may be associated with DFS and OS in young women with breast cancer. Further validation and feasibility studies are required before the result of our study can be considered for clinical practice.

Prognostic value of circulating microRNA-21 in digestive system cancers: a meta-analysis.

UNLABELLED: Circulating microRNAs show aberrant expression in patients with cancer. The aim of this study was to investigate the prognostic value of circulating microRNA-21 (miR-21) in digestive system cancers. METHODS: All the eligible studies were searched by Medline and EMBASE. The hazard ratios (HRs) for overall survival (OS), which compared the expression levels of circulating miR-21 in patients with digestive cancer was extracted and estimated. Pooled HRs and 95% confidence intervals (CI) were calculated. Then a meta-analysis was performed to clarify the prognostic value of the miR-21. RESULTS: A total of seven studies involving 907 subjects were included. The results suggested that higher circulating miR-21 could predict worse OS outcome with the pooled HR of 2.19 (95% CI 1.01-4.75, P = 0.05) in digestive system cancers. Subgroup analysis by ethnicity indicated circulating miR-21 was associated with OS in patients with digestive cancer among Asians with the pooled HR of 2.90 (95% CI 1.30-6.45, P = 0.009). However, subgroup analysis by digestive system site revealed that there is no associated with OS in patients with colorectal cancer with the pooled HR of 1.34 (95% CI 0.45-4.00, P = 0.60). CONCLUSION: The present findings suggest that circulating miR-21 is associated with poor survival in patients with digestive cancer and could be a prognostic biomarker for those patients.

Thymectomy in central lymph node dissection for papillary thyroid cancer.

BACKGROUND: Central lymph node dissection (CND) has been proposed in the treatment of patients affected by papillary thyroid cancer (PTC) with clinically negative neck lymph nodes. The procedure allows pathologic staging of lymph nodes of the central compartment and treatment of the micrometastases. By comparing bilateral and unilateral thymectomy during total thyroidectomy with central lymph node dissection for postoperative complications in sonographically node-negative papillary thyroid carcinomas, we aimed to determine the optimal extent of prophylactic central lymph node dissection. METHODS: Patients were divided into two study groups: Group 1, total thyroidectomy plus unilateral thymectomy during the CND; Group 2, total thyroidectomy associated with bilateral thymectomy (both upper poles) during the CND. Primary endpoints of the study were evaluated by comparing the postoperative complications between the two groups. RESULTS: The only significant result found when comparing the two groups was the rate of transient hypocalcemia. (Group 1: 13.7%, Group 2: 52.4%, p<0.01). A total of five cases of papillary thymic metastases were found in this study. And final pathology confirmed that all cases of thymic metastases were lymph node micrometastases of PTC, only situated in the ipsilateral thymus upper pole. CONCLUSIONS: Bilateral thymectomy during the CND did not provide a better carcinologic resection, as no contralateral thymic metastases were found. The unilateral thymectomy with total thyroidectomy during the CND may represent an effective strategy for reducing the rate of postoperative hypocalcemia.

Continued twitching of the latissimus dorsi miniflap after breast conservation therapy: a case report.

We report a case of continued twitching of the latissimus dorsi muscle following breast conservation therapy, along with immediate reconstruction with a latissimus dorsi miniflap, which continued despite several attempts at control including BTX-A percutaneous local injection, and was finally cured by delayed division of the thoracodorsal nerve via a small well-tolerated axillary incision.

Prognostic and metastatic value of phosphatase of regenerating liver-3 in invasive breast cancer.

PURPOSE: The aim of this study was to investigate the expression of the PRL-3 in human invasive breast cancer and to evaluate its clinical and prognostic significance. Its potential role in the invasive-metastatic properties of invasive breast cancer was also investigated. METHODS: Protein expression of PRL-3 was evaluated by immunohistochemistry for a consecutive series of 82 invasive human breast cancer tissues and 63 matched lymph node metastases, including PRL-3 mRNA expression analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) in malignant, nonmalignant breast tissue samples and lymph node metastases. We investigated the correlation of PRL-3 with clinicopathologic features, Overall and recurrence-free survival distribution curves were assessed using the Kaplan-Meier test and log-rank statistics, followed by Cox proportional hazards regression model. RESULTS: We found that 70.7% patients expressed a high level of PRL-3 protein in their tumors, and its over expression was positive correlated with lymph node metastasis (LNM) (P = 0.011). Moreover, The PRL-3 mRNA expression was significantly higher in malignant compared to benign breast tissue, while increased expression of PRL-3 mRNA was significantly associated with LNM (P = 0.002). Univariate analysis showed that the positive expression of PRL-3 was a poor risk prognostic factor (OS, P = 0.045; RFS, P = 0.034). Multivariate analysis using the Cox regression model indicated that high PRL-3 expression was an independent unfavorable prognostic factor for RFS. CONCLUSIONS: These results strongly suggest that PRL-3 expression can indicate the potential role of LNM to some extent. Increasing the risk of tumor metastasis (OR = 3.889). Our results also imply that PRL-3 might be a novel molecular marker for predicting relapse of invasive breast cancer.

[Laparoscopic abdominoperineal resection for low rectal cancer].

OBJECTIVE: To assess the advantage and disadvantage of laparoscopic abdomino-perineal resection and open abdominoperineal resection for low rectal cancer. METHODS: Patients with low rectal cancer, collected from July 2003 to April 2006, were randomly divided into laparoscopic abdominoperineal resection group (37 cases) and open abdominoperineal resection group (37 cases). Operation time, number of lymph node removed, intra-operative blood loss, time to pass flatus, time to ambulate, time to discharge, complications, early recurrence, and economical cost were compared between the 2 groups. RESULTS: All patients were performed successfully. For the first 10 patients, operation time of laparoscopic group was significantly longer than that of open group, but there was no significant difference between the 2 groups. Intra-operative blood loss of laparoscopic group was significantly less than that of open group, but it was reverse for the first 10 patients. There was no significant difference in time to pass flatus between the 2 groups. Time to ambulate in laparoscopic group was significantly earlier than that in open group. There was no significant difference in time to discharge between the 2 groups, but it was earlier for perineum closure in laparoscopic group. Relative complications of laparoscopic group, including pulmonary infection, abdominal wound infection or split, were significantly less than those of open group. There was no significant difference in number of lymph nodes removed, early recurrence between the 2 groups. Operation cost of laparoscopic group was significantly higher than that of open group, but there was no significant difference. CONCLUSION: Advantages of laparoscopic abdominoperineal resection were characterized for not only minimal invasion and good cosmetic outcome but also less blood loss, complications, and earlier postoperative recovery. The operation time, total costs and oncological clearance of laparoscopic abdominoperineal resection patients were comparable with those of open procedure patients.