Application Principle Of Laparoscopic Ultrasound Guided Radiofrequency Ablation Of Liver Cancer And Research On Effect Of MiR-130a On Migration And Invasion Of Liver Cancer

Objective:1. To summarize application principle of laparoscopic ultrasound-guided radiofrequency ablation (LRFA) of liver cancer, and to analyze the application characteristics, causes and prevention of complications and prognosis.2. To discussion the significance of three-dimensional reconstruction as a method of preoperative planning of LRFA.3.To find out miR-130a expression in hepatocellular carcinoma of liver cancer and the impact of migration and invasion.Methods:1.905 cases of LRFA from Jan 2006 to Dec 2015 in our center were retrospectively studied. The clinical data, surgical procedure, post-surgical information and survival time were collected in order to analyze the application characteristics, causes and prevention of complications and prognosis.2.32 cases of LRFA in recent 24 months in our center were analyzed for which three-dimensional reconstruction were taken as a method of preoperative planning, with pairing other 64 cases of LRFA as a case-control study. Evaluate the significance of three-dimensional reconstruction as a method of preoperative planning of LRFA.3. With method of qRT-PCT, inhibitor vector down-regulation, Transwell、Western Blotting, to find out miR-130a expression in hepatocellular carcinoma of liver cancer and the impact of migration and invasion.Result:Chapter 1:1. LRFA is an increasingly important part of the comprehensive treatment of liver cancer, the number of cases increased year by year. There were few cases took LRFA as first treatment of liver cancer (P<0.05), more with Child-Pugh B or C liver function (P<0.05) and more with tumor diameter>30mm (P<0.05) in cases of 2006-2010. Contrast to percutaneous approach, LRFA is more suitable for tumor is: ① Localization of the HCC within 1 cm of the liver capsule or extrahepatic protrusive HCC. ② Localization in the dome of the liver (difficult needle placement in axial plane).③ Adjacent to the gastrointestinal tract, gallbladder, bile duct or heart.2. We use ablation zone in Enhanced Imaging of one month after LRFA to determine whether it’s complete response.89.72%(812) cases were visit one month after LRFA with a total ablation rate of 87.31%. Ablation rate of 2006-2010 and 2011-2015 cases were 81.41% and 92.17% respectively, P< 0.001. The results of logistic regression analysis found that profound predictors for complete response rate of LRFA were as follows: ① dangerous location(OR= 3.225, P= 0.000), ② cirrhosis (OR= 1.110, P= 0.040), ③ liver function Child-Pugh rating (P= 0.000), ④ tumor size (P= 0.000), ⑤ tumor number (P= 0.001).3. Biopsy ratio and positive biopsy rate of the two groups (53.62% and 74.82%) are all relatively low. Cases of 2006-2010 had more post-operative stay (P<0.001) and post-operative ALT changes (P=0.011)4. In aspect of incidence of complications, there’s no significant difference of Clavien-Dindo Ⅰ-Ⅱ complications, but 2011-2015 cases has low incidence of Clavien-Dindo Ⅲ and Ⅳ complications (P=0.033、0.029). Because LRFA is performed under direct vision, the incidence of abdominal bleeding (18/905), pneumothorax (15/905), bile leakage (4/905) is low. Tumor metastasis (4/905) should be avoided by reducing the number of puncture and increasing protective measures.5.576 patients from 2011 to 2015 participated in return visit for more than 3 months after the LRFA, range 3-60 months. Median disease free survival (median DFS) was 23.0 months. The overall survival (OS) rate of 12,24 and 36 month were 38.24%, 50.87% and 52.44% respectively. COX proportional hazards regression analysis indicated that significant predictors for DFS after LRFA of 2011-2015 patients were as follows:① dangerous location (RR= 2.130, P= 0.012),②metastatic or recurrent HCC (RR= 3.488, P= 0.003), ③ liver function Child-Pugh rating (P= 0.000), ④ tumor size (P= 0.000), ⑤ tumor number (P= 0.001),⑥ the preoperative presence of obstructive jaundice (RR= 1.633, P= 0.031), ⑦ the preoperative presence of portal vein thrombosis (RR= 2.311, P= 0.005) ⑧ diabetes (RR= 1.168, P= 0.028).Chapter 2:1.32 cases of LRFA in recent 24 months in our center were analyzed for which three-dimensional reconstruction were taken as a method of preoperative planning, with pairing other 64 cases of LRFA as a case-control study. Compared with LRFA group,3D-RFA group has shorter operation time(p=0.0023) and shorter mean puncture time (P=0.0214). There’s no significant difference of blood loss (P=0.1813)2. There’s no significant difference of ablation rate and incidence of complications (p=0.871).3D-RFA group has shorter post-operative stay (P=0.0160) and post-operative TNF-a changes (0.0273)3. DFS between the two groups was significantly different (P=0.0442). DFS of 12 months survival rates was 77.636% and 65.736% respectively. Median DFS of was LRFA group was 16 months and 3D-LRFA group over 24 months.4. The standard route of LRFA is established by concluding preoperative planning with three-dimensional reconstruction. The standard route of LRFA will significantly shorten the learning curve of surgeons.Chapter 3:1. Expression of miR-130a in hepatocellular carcinoma was significantly higher than the corresponding adjacent tissues (P=0,001). The miR-130a expression in liver cancer cell lines was significantly higher than in normal liver cells (P= 0,000).2. Transwell results showed that, compared with control cells, migration and invasion of highly metastatic liver cancer cell lines (MHCC97-H、HCCLM3) are significantly influenced by reducing the level of miR-130a with transfecting with miR-130a inhibitor (P=0.0004,0.0001,0.0002,0.0005)3. Western Blotting results showed that, expression of MMP-9 and MMP-2 are significantly decreased in miR-130a downregulated MHCC97-H, HCCLM3 cell lines (P=0.003、0.004、0.000、0.006). This study demonstrated that miR-130a promotes the process of EMT in liver cancer cells by certain signaling pathways and molecular mechanisms.Conclusion:1:With advantage of LUS and laparoscopic surgery, as a useful complement to ultrasound-guided percutaneous ablation of liver cancer, LRFA of liver cancer can reduce the incidence of postoperative complications and increase the safety of operation. It reflects concept of accurate, minimally invasive treatment and a good clinical value. For patients with small unresectable hepatocellular carcinoma or tumor located in dangerous place, LRFA is a good and promising surgical treatment, especially for patients with cirrhosis. But LRFA still need high skills of laparoscopic liver surgery and laparoscopic ultrasound technology.2. Three-dimensional model of liver reconstruction based on image information is a powerful tool of iver surgery planning. It helps to simulate tumor location and vital tubular structure, make plan for interventional treatment, and therefore mean puncture time and operation time is shortened, influence on liver function is reduced, hospital stay is decreased and DFS is prolonged. Three-dimensional model of liver reconstruction is important complement for LRFA.3. miR-130a facilitates the process of EMT in liver cancer cells by certain signaling pathways and molecular mechanisms, resulting in increased migration and invasion of liver cancer cells. The target genes and the corresponding signal pathways of miR-130a are still worthy of further study. miR-130a may become one of the molecular therapy target of liver cancer.