Alternative management of anatomical right hemihepatectomy using ligation of inflow and outflow vessels without hilus dissection.

BACKGROUND AND AIM: The conventional method of anatomical right hemihepatectomy (ARHH) requires hilus dissection. We report a method without hilus dissection to minimize intraoperative bleeding. METHODS: We retrospectively evaluated data of 107 patients who received ARHH involving ligation of corresponding inflow and outflow vessels (LCIOV) without hilus dissection between January 2000 and October 2008. Results were compared to those of patients who underwent non-anatomical right hemihepatectomies (NARHH). RESULTS: The two groups had similar gender and age (both, P>0.05). The LCIOV group had a higher percentage of patients without intrahepatic metastases (94.6% vs 80.3%, P=0.003). Hepatocellular carcinoma (HCC) lesion size (9.3 vs 10.2, P=0.023), durations of inferior vena cava occlusion (4 vs 4.7, P<0.001) and portal triad occlusion (7 vs 11, P<0.001), blood loss (430 vs 580 mL, P=0.001), transfusion volume (300 vs 520 mL, P<0.001), and measures of postoperative liver function (e.g. maximum aspartate aminotransferase [AST]) of the LCIOV group were also significantly less than the NARHH group. Larger hepatic cavernous hemangiomas (HCH) lesion size (16.2 vs 13.0, P<0.001), longer operative time (168 vs 154 min, P=0.017), and a lower percentage of patients with inferior vena cava occlusion (17.8% vs 35.2%, P=0.001), pleural effusions (19.3% vs 30.9%, P=0.042), and blood transfusions (10.3% vs 75.0%, P<0.001) were found in the LCIOV group. CONCLUSION: The reported method is a safe and bloodless technique for right hemihepatectomy in select patients.

A new simple and safe technique of end-to-end invaginated pancreaticojejunostomy with transpancreatic U-sutures--early postoperative outcomes in consecutive 88 cases.

BACKGROUND: Postoperative pancreatic fistula (POPF) remains a leading cause of morbidity and mortality after pancreaticoduodenectomy (PD). Thus, a number of technical modifications regarding the pancreato-enteric anastomosis after PD have been proposed to reduce POPF rate. Until now, there is no consensus on which is the best. This study presents a new technique of the end-to-end invaginated pancreaticojejunostomy with two to three transpancreatic U-sutures and evaluates its safety and reliability. MATERIAL AND METHODS: From 2002 to 2007, 88 patients (54 men and 34 women) underwent an invaginated end-to-end pancreaticojejunostomy with two to three transpancreatic U-sutures after PD. The mean age was 52.4 years (range, 26-74 years). The diseases of the all patients were malignant. RESULTS: In all patients of this study, two transpancreatic U-sutures were performed in 59 and three U-sutures in 29. The median duration of surgery was 3.8 h (range 3-6.5) and the median time to perform pancreaticojejunostomy was 13.3 min (range 8-25). The median blood loss was 750 ml (range 300-1,800), 36 patients needed transfusion and the median blood transfusion was 380 mL (range 200-1,200). Overall morbidity occurred in 15 patients (17.0%). Only two patients (2.2%) had grade A of POPF and no patient had grade B and grade C of POPF. No operative death occurred. CONCLUSIONS: An invaginated end-to-end pancreaticojejunostomy with two to three transpancreatic U-sutures is simple, rapid, safe, and reliable technique, even in some patients with soft pancreas and small pancreatic duct.

Right hepatectomy using the liver double-hanging maneuver through the retrohepatic avascular tunnel on the right of the inferior vena cava.

BACKGROUND: The key to Belghiti's liver-hanging maneuver is to develop a retrohepatic tunnel. This procedure requires a blind dissection of the plane anterior to the inferior vena cava (IVC), with the inherent risks of damaging the short hepatic veins and consequential bleeding. The aim of this article is to describe a liver double-hanging maneuver with the advantage of being technically simple and safe. METHODS: The operator uses his or her right index finger to dissect the space from below upward between the hepatic parenchyma and the anterior and superior edge of the right adrenal gland, which is situated just on the right side of the IVC. The operator then uses his left index finger to dissect the retrohepatic space from above downward on the right side of suprahepatic IVC, which is lateral to where the right hepatic vein joins the IVC. The retrohepatic tunnel is built when the 2 fingers touch each other. A kidney pedicle forceps is used to place 2 tapes around the liver for suspension. RESULTS: In all, 65 patients underwent right hepatectomy using this maneuver. The study included 62 patients with hepatocellular carcinoma (tumor size: mean +/- SD, 10 +/- 3.7 cm), and 3 patients had hepatic cavernous hemangioma, with a maximum diameter of 12.6 cm, 14.4 cm, and 22.6 cm, respectively. No major bleeding was encountered during the creation of the retrohepatic tunnel, with a success rate of 100%. CONCLUSION: To develop the retrohepatic tunnel in the space on the right of the IVC is absolutely bloodless, and it is technically easy and safe.

[The technique of radical pancreaticoduodenectomy for malignant tumor in pancreatic head with pressed superior mesenteric blood vessels or portal vein].

OBJECTIVE: To investigate the technique of radical pancreaticoduodenectomy for malignant tumor in pancreatic head with pressed superior mesenteric blood vessel or portal vein. METHODS: From March 2005 to March 2007, thin slice scan and vessel-reconstruction of 56 patients of malignant tumor in pancreatic head with pressed superior mesenteric blood vessels or portal vein were carried out using multidetector spiral CT to evaluate whether peripheral vessels of pancreatic tumor were invaded and whether the tumor was resectable. During the operation, 3 vascular blocking bands for superior mesenteric vein, portal vein and spleen vein or 4 vascular blocking bands (additional one for inferior mesenteric vein) were preset. Under the cross and traction between superior mesenteric vein and superior mesenteric artery, resected the uncinate process of pancreas thoroughly. Using those methods, radical pancreaticoduodenectomy for 56 patients above-mentioned were successfully accomplished. RESULTS: The accuracy for preoperative judging by using multidetector spiral CT whether the peripheral vessels of pancreatic cancer were invaded and whether the tumor was resectable was 98% and 100% separately. Thirty-seven of 56 patients, whose superior mesenteric blood vessels or portal veins were pressed by the tumor of pancreatic head, were operated using 3 vascular blocking bands and 2 patients using 4 vascular blocking bands, followed by suturing the bleeding points of the superior mesenteric vein with 5-0 vascular suture Proline. One patient's superior mesenteric vein was partially resected and restored. The operations cost 5-8 h each and the blood loss was 200-600 ml. There were no operative or postoperative hemorrhage or pancreatic juice leakage. According to the follow-up up to now, 2 patients died of multiple live tumor metastases 7 and 9 months separately after operation, the other 54 patients were still alive. CONCLUSIONS: Thin slice scan and vessel-reconstruction using multidetector spiral CT can accurately judge whether the blood vessels near the pancreatic tumor were invaded and whether the tumor was resectable, using 3 vascular blocking bands or 4 vascular blocking bands and cross, traction of the superior mesenteric blood vessels, operator can easily accomplish the radical pancreaticoduodenectomy of malignant tumor in pancreatic head with pressed superior mesenteric blood vessels and portal vein, which was not resectable or need combined resection of the blood vessels in the traditional opinion.

Mesohepatectomy for hepatocellular carcinoma: a study of 256 patients.

BACKGROUND: Mesohepatectomy is a rarely used operative procedure to treat liver cancer because of its technical complexity. In patients with hepatocellular carcinoma (HCC) with a viral hepatitic/cirrhotic background, this procedure can be used to resect the tumor with adequate margins, while at the same time preserve as much functioning hepatic parenchyma as possible. This retrospective study aimed to evaluate the early results and late survival outcomes of mesohepatectomy in HCC. MATERIALS AND METHODS: From 1996 to 2005, 256 patients with HCC situated at the central liver segments (Couinaud segments IV, V, VIII+/-I) were treated with mesohepatectomy. The treatment outcomes of these patients were retrospectively analyzed. RESULTS: The in-hospital mortality rate was 0.4%, but the postoperative morbidity rate was 28.1%. The 1-, 3-, and 5-year overall survival rates were 77.0, 49.8, and 35.1%, while the 1-, 3-, and 5-year disease-free survival rates were 59.1, 28.8, and 17.0%, respectively. Multivariate analyses showed the significant factors for overall survival were tumor size>8 cm, vascular invasion, and alpha fetoprotein (AFP)>5,000 ng/ml; and for disease-free survival were tumor size>8 cm, vascular invasion, tumor number (three or more), AFP>5,000 ng/ml. CONCLUSION: Mesohepatectomy is a safe and effective treatment for a centrally situated HCC with a viral hepatitic/cirrhotic background.

A simple technique ligating the corresponding inflow and outflow vessels during anatomical left hepatectomy.

BACKGROUND: Massive bleeding remains one of main factors of morbidity and death in liver resections. For this reason, the Pringle maneuver or total vascular exclusion is commonly used during liver resection. However, ischemic damage is still a major problem. Some surgeons used the "glissonean" approach for bleeding control, but the technique is tedious and also time consuming, with high incidence of bile leaks in the postoperative period. The aim of this paper is to describe a new bleeding control technique, rapid ligation of the corresponding inflow and outflow vessels without hilus dissection before the parenchyma transection during anatomical left liver resection and to analyze the feasibility, blood loss, transfusion requirements, and postoperative complications. MATERIALS AND METHODS: During the past 18 years, we used the new hemorrhage control technique in left liver resection in 630 patients with malignant or benign tumors. RESULTS: The median blood loss in all 630 patients was 110 +/- 250 ml (range 50-750), and no patient required blood transfusion. The median total operative time was 77 +/- 35 min (range 25-155). No bile leaks and liver failure of the patients occurred postoperatively. There was no death within 30 postoperative days. CONCLUSION: The rapid ligation of the corresponding inflow and outflow vessels without hilus dissection before the parenchyma transection is a feasible, safe, and bloodless technique during the left liver resection.

[Effects of mitofusin-2 gene on proliferation and chemosensitivity of human breast carcinoma cell line MCF-7].

BACKGROUND & OBJECTIVE: Mitofusin-2(mfn2), a proliferation-inhibiting gene, targets to the outer membrane of mitochondria. Its overexpression suppresses the proliferation of vascular smooth muscle cells. This study was to explore the effects of mfn2 gene on the proliferation and chemosensitivity of human breast carcinoma cell line MCF-7. METHODS: Plasmid pEGFP-mfn2 containing mfn2 cDNA was constructed and transfected into MCF-7 cells by sofast. The expression of green fluorescent protein (GFP) in MCF-7 cells was detected by Western blot. Cell proliferation was measured by MTT assay and cell counting. Cell cycle and chemosensitivity of MCF-7 cells to camptothecin (CAM) was observed by flow cytometry (FCM). RESULTS: After transfection of pEGFP-mfn2, the stable expression of GFP protein was detected in MCF-7 cells, and cell cycle was arrested: the S phase proportion was significantly higher in pEGFP-mfn2-transfected cells than in pEGFP-transfected and untransfected cells [(42.7+/-1.3)% vs. (17.2+/-2.0)% and (19.6+/-1.7)%, P<0.05]. The apoptosis rate were significantly higher in pEGFP-mfn2-transfected cells than in pEGFP-transfected and untransfected cells [(16.0+/-0.3)% vs. (4.5+/-0.9)% and (3.6+/-0.6)% before treatment of CAM, P<0.05; (69.6+/-4.3)% vs. (31.0+/-1.8)% and (23.4+/-2.8)% after 4-hour treatment of CAM, P<0.05]. CONCLUSION: mfn2 gene can inhibit the proliferation of MCF-7 cells and increase their chemosensitivity to CAM.

Role of mesohepatectomy with or without transcatheter arterial chemoembolization for large centrally located hepatocellular carcinoma.

BACKGROUND: The role of preoperative transcatheter arterial chemoembolization (TACE) for resectable hepatocellular carcinoma (HCC) was controversial. METHODS: 246 patients with large centrally located HCC underwent mesohepatectomy (MH) and were divided into two groups: group A, 89 patients with preoperative TACE; group B, 157 patients without preoperative TACE. The aim was to evaluate the influence of preoperative TACE on postoperative complications and long-term results of patients with large centrally located HCC. RESULTS: In the 89 patients of the TACE-MH group, a total of 123 (mean 1.4 per patient) preoperative TACEs were performed. The differences in postoperative complications (34.8 vs. 24.2%; p=0.075) and overall hospital mortality (3.4 vs. 0.6%; p=0.103) between the two groups were not significant. The postoperative recurrence rate in the remnant liver was higher in the MH group than in the TACE-MH group (79.6 vs. 73.0%), while the extrahepatic metastasis rate in the TACE-MH group was higher than that in the MH group (11.1 vs. 7.0%). Overall 1-, 3-, and 5-year survival rates were 87.1, 62.9, and 46.2%, respectively, for the TACE-MH group, and 82.2, 54.4, and 31.7%, respectively, for the MH group (p=0.001); 1-, 3-, and 5-year disease-free survival rates were 75.0, 46.2, and 31.8%, respectively, for the TACE-MH group, and 69.6, 38.0, and 16.5%, respectively, for the MH group (p=0.002). CONCLUSIONS: Long-term outcomes of patients with preoperative TACE were improved and the pattern of the recurrences after surgery was altered. The patients with large centrally located HCC could benefit more from this neoadjuvant treatment, although there was some influence of preoperative TACE on postoperative complications.

[Significances of gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time].

OBJECTIVE: To investigate the gene differential expression patterns in hepatocirrhosis and non-hepatocirrhosis tissues within different ischemic time. METHODS: The liver tissues were divided into two groups: Group A (non-hepatocirrhosis), Group B (hepatocirrhosis), each of which consisted of 3 groups with different ischemic time: 15, 30 and 45 minutes. The gene differential expression patterns in the two groups within different ischemic time were detected and compared with those in normal liver tissues by using 4000 points gene microarray. RESULTS: In non-hepatocirrhosis tissues, the homeostatic maintenance genes expressed highly during hepatic ischemia for 15 minutes, and no apoptotic gene was expressed; but in hepatocirrhosis tissues, many apoptotic genes expressed highly. As for 30 minutes, in both two groups liver tissue genes expressed to the peak, and the genes related to cell death, oxidative stress and nuclear factors expressed highly. The difference lies in the facts that in Group B pro-apoptosis genes expressed more than those in Group A, and the Ratio values were higher than those in Group A. Many genes of heat shock protein family and antioxidant proteins expressed highly simultaneously in Group A, but comparatively low in Group B. As for 45 minutes, genes of heat shock proteins and antioxidant proteins expressed lowly in Group B. CONCLUSIONS: It suggests that the safe time limit of hepatic ischemia for cell survive is 30 minutes or so. Non-hepatocirrhosis tissues could endure 30 minutes of ischemia and even longer, but it should be restricted within 30 minutes in hepatocirrhosis tissues.

[Effect of mitofusin-2 gene in apoptosis of human breast carcinoma cell line in vitro].

OBJECTIVE: To investigate the role of mitofusin-2 gene (mfn2) in apoptosis in human breast carcinoma cell line MCF-7 cells after in vitro transfection. METHODS: pEGFP mfn2 was transfected by sofast in vitro. Expression of GFP was observed by Western blot, and the MCF-7 cell proliferation was measured by MTT and cell counting. Apoptosis in MCF-7 cells was observed in annexin-V/PI and chondrosome transmembrane potential of MCF-7 marked in JC-1 by FCM. The Ultrastructure of cells was observed by transmission electron microscopy. RESULTS: The stable expression of GFP in MCF-7 cells was confirmed by Western blot. Mfn2 significantly inhibited cell proliferation, revealed by MTT, and decrease chondrosome transmembrane potential. Exogenous mfn2 gene significantly induced apoptosis. The apoptotic rate was increased from 3.6% to 16.0% (P < 0.05). Mfn2 gene induced break down and loss of mitochondrial cristae, and rarefaction of mitochondrial ground substance. Swollen mitochondria intensely aggregated around the cell nuclei. CONCLUSION: Mfn2 can strongly induce apoptosis in MCF-7 cells, which may be associated with decrease of mitochondrial transmembrane potential.

Hepatectomy for huge hepatocellular carcinoma in 634 cases.

AIM: To clarify the safety and feasibility of hepatectomy for huge hepatocellular carcinoma (HCC). METHODS: A total of 4765 patients with HCC operated at Tongji Hospital were retrospectively studied, of them, 780 patients had huge HCC (10 cm or more in diameter). Hepatectomy was carried out on 634 patients (81.2%). The majority of the liver resection were major resections, and combined resection of the adjacent organs or structures was common (17.2%). The liver resection was combined with portal vein thrombectomy in 139 patients (21.9%). RESULTS: Postoperative complications were common (26.8%) and required another laparotomy to prevent the complications in 5 patients (0.8%). The 30-d mortality was 2.2%. The main causes of postoperative deaths were liver failure (n = 9), postoperative bleeding (n = 4) and septic complication (n = 1). The 3-, 5- and 10-year survival rates after liver resection were 35.1%, 18.2% and 3.5%, respectively. CONCLUSION: Hepatectomy for huge HCC is safe and effective. It should be used to treat patients with low surgical risks and resectable tumours.

Effects of location and extension of portal vein tumor thrombus on long-term outcomes of surgical treatment for hepatocellular carcinoma.

BACKGROUND: The role of surgical resection and thrombectomy for hepatocellular carcinoma (HCC) with portal vein tumor thrombus (PVTT) is controversial. This study aimed to evaluate the effects of the location and extent of PVTT on the long-term outcomes of surgical treatment for HCC. METHODS: A total of 438 patients with HCC and PVTT underwent liver resection with or without thrombectomy. These 438 patients were divided into 2 groups: in group A, PVTT was located in the hepatic resection area or protruded into the first branch of the main portal vein beyond the resection line for < 1 cm (286 patients), and in group B, PVTT extended into the main portal vein (152 patients). Concomitant thrombectomy was performed in 147 patients (51.4%) of group A and in all patients of group B. RESULTS: PVTT recurrence within 6 months after surgery in group B was significantly higher than that in group A: 76.9% vs. 11.3%. Remnant liver recurrence within 1 year after surgery was 45.0% in group A and 78.8% in group B. The cumulative 1-, 2-, 3-, and 5-year overall survival rates were 58.7%, 39.9%, 22.7%, and 18.1% for group A and 39.5%, 20.4%, 5.7%, and 0% for group B, respectively. The overall survivals were significantly better in group A than group B (P < .02). CONCLUSIONS: Liver resection with thrombectomy yielded better outcomes in the HCC patients with PVTT confined to the first or second branch of the main portal vein compared with PVTT extending into the main portal vein.

Modified technique of hepatic vascular exclusion: effect on blood loss during complex mesohepatectomy in hepatocellular carcinoma patients with cirrhosis.

BACKGROUND: Hepatic veins remain patent during complete inflow occlusion (CIO) and bleeding from them may continue. Occlusion of the inferior vena cava (ICV) during CIO may reduce blood loss from hepatic veins. This study was designed to compare the overall outcomes after application of CIO with or without occlusion of the ICV below the liver in complex mesohepatectomy for hepatocellular carcinoma (HCC) patients with cirrhosis. MATERIALS AND METHODS: One hundred and eighteen (118) patients were randomly assigned to CIO or a modified technique of hepatic vascular exclusion (MTHVE). Hemodynamic parameters were evaluated and the amount of blood loss, measurement of liver enzymes, and postoperative progress were recorded. RESULTS: Blood loss during liver transection in CIO groups was significantly greater than that in MTHVE group (P=0.046). Thus, incidence of blood transfusion was significantly greater in patients of the CIO group (P=0.041). There were no significant differences in liver enzyme changes, bilirubin, or morbidity in the postoperative period between the two groups. CONCLUSIONS: CIO with occlusion of the ICV below the liver is a safe, effective, and feasible technique during mesohepatectomy in HCC patients with cirrhosis. Excellent results were obtained with minimized bleeding, limited hepatic function damage, and low rate of postoperative complications.

[Study of immunological effect of dendritic cell transfected with survivin gene on the specific anti-alimentary tract tumor].

OBJECTIVE: To investigate the effects of dendritic cells (DCs) transfected with survivin gene, and to observe the effective and specific anti-tumor immunological effect induced by modified DC in vitro. METHODS: Survivin gene was transfected to DCs with liposomes. Survivin expression could be detected both in DCs cells and in cell culture with method of Western blot. Cytokines as well as cellular surface molecule such as IL-12, TNF-alpha, CD1 alpha, CD83, MHCII, CD80 and CD86 were detected. The competence of inducing human specific cytotoxic T lymphocyte (CTLs) was also detected with MTT. RESULTS: Survivin expression could be detected both in DCs which were transfected with survivin cDNA and in cell culture superior. The IL-12 and TNF-alpha level was (265.2 +/- 32.7), (437.1 +/- 83.5) pg/ml, and much higher in transgened DC cells than blank DC cells (P < 0.05). CD1 alpha, CD83, MHCII, CD80 and CD86 was high expressed in survivin-DC cells, however, it was low expressed in blank DC cells. The lyse rate to gastric cancer cell, colon cancer cell and bile duct cancer cell was 65%, 77%, and 85% respectively, and these were much higher than those of blank DC cells. CONCLUSIONS: DCs transfected with survivin gene could induce specific cytotoxic T lymphocytes and strikingly raised DC cell's antigen present function, and have specific CTL killing activity.

[Role of united hepatectomy and splenectomy in the surgical treatment of hepatocellular carcinoma complicated with hepatic cirrhosis and hypersplenism].

OBJECTIVE: To investigate the role of united hepatectomy and splenectomy in the surgical treatment of hepatocellular carcinoma complicated with hepatic cirrhosis and hypersplenism. METHODS: Two hundred and four patients of hepatocellular carcinoma complicated with liver cirrhosis and hypersplenism were divided into two groups: the group of combined resection of hepatocellular carcinoma and spleen (group A, n = 94) and the group of hepatectomy only (group B, n = 110). The counts of white blood cell and platelet, total serum bilirubin levels, changes of immune function, operative morbidity and 5-year survival rates were compared between the two groups. RESULTS: (1) There was no significant difference of the counts of CD4, CD8, CD4/CD8 and the levels of IL-2, IFN-gamma and IL-10 between the two groups before the operation. (2) Two months after operation, the percentage of CD4 and the ratio of CD4/CD8 were significantly higher in the group A [(40.8 +/- 4.1)% and (1.8 +/- 0.2)%, respectively] than those of group B [(33.8 +/- 3.6)% and (1.1 +/- 0.3)%, respectively], while the percentage of CD8 was (25.8 +/- 3.8)% in the group A, significantly lower than that of group B [(32.9 +/- 4.1)%, P < 0.05]; Both the levels of IFN-gamma and IL-2 were significantly higher in the group A than those of group B while the level of IL-10 in group A was lower compared with that of group B (P < 0.05). (3) On the 14 postoperative day, the counts of white blood cell and platelet were (9.1 +/- 1.4) x 10(9)/L and (310 +/- 55) x 10(9)/L, which were significantly higher than those of group B [(3.6 +/- 1.2) x 10(9)/L and (99 +/- 36) x 10(9)/L, respectively]. (4) On the 7th postoperative day, the total serum bilirubin concentration of group A [(24 +/- 7) micromol/L] was lower than that of group B [(37 +/- 13) micromol/L]. (5) There was no significant difference in the postoperative morbidities between the two groups (15.9% and 14.5%, respectively). (6) There was no significant difference of the 5-year cumulative survival rates between group A (56.4%) and group B (50.9%, P > 0.05), but the survival rate without tumor of group A was 37.7%, higher than that of group B (18.9%, P < 0.05). CONCLUSIONS: The combined resection of hepatocellular carcinoma and spleen for the hepatocellular carcinoma complicated with liver cirrhosis and portal hypertension may promote the recovery of the balance between the subgroup of T cell and B cell, normalize the counts of white blood cell and platelet, alleviate the bilirubin burden and benefit for the recovery of liver physiological role without increase; the 5-year disease-free survival rate was improved significantly while no increase of postoperative morbidity. Combined resection may also be helpful for the delay of the progression of liver cirrhosis and for the prevention of esophageal variceal bleeding.

[A candidate tumor suppressor gene mutated in primary hepatocellular carcinoma: kruppel-like factor 6].

OBJECTIVE: To explore the relationship between kruppel-like factor (KLF)6 gene and the development or progression of hepatocellular carcinoma (HCC). METHODS: Reverse-transcription polymerase chain reaction (RT-PCR) was used to examine the expression of KLF6 mRNA in normal liver tissue and primary hepatocellular carcinoma, and single strand conformation polymorphism (SSCP), DNA sequencing were used to detect the point mutation of KLF6 in primary hepatocellular carcinoma. RESULTS: An amplified fragment of 427 bp DNA was detected in 31 (97%) of 32 adjacent noncancerous tissue and normal liver tissue, and in 23 (85%) of 27 HCCs. There was no significant difference in the levels of KLF6 mRNA between normal liver and liver tumors (chi(2) = 2.58, P > 0.05). For the 27 HCCs, six SSCP-positive bands (22%) were detected. Among them, three of 5 (3/5) tumor samples showing loss of heterozygosity (LOH) of KLF6 had mutations in the retained KLF6 allele. CONCLUSION: We showed that LOH was detected in 5 (36%) HCCs obtained from 14 informative cases, and three of 5 tumor samples showing LOH of KLF6 had mutations in the retained KLF6 allele. Two inactivating events had occurred; thus, as defined by Knudson's "two-hit model", 16 KLF6 appears to be a tumor suppressor gene.

Effect of octreotide on human pancreatic cancer cells after transfected with somatostatin receptor type 2 gene.

AIM: To observe the effect of octreotide on apoptosis rate of human pancreatic cancer cells PC-3 after transfected with somatostatin receptor type 2 (SST2) gene. METHODS: SST2 plasmid was transfected into PC-3 cells by liposome. Result of transfection was detected by immunocytochemical staining and Western blotting. Apoptosis rates of PC-3 cells under different dosages of octreotide were measured by MTT assay and flow cytometry (FCM). RESULTS: Apoptosis rate caused by octreotide of transfected PC-3 cells was 7.56+/-1.06% at the dosage of 0.20 microg/mL, 9.25+/-1.73% at the dosage of 0.40 microg/mL and 14.18+/-2.71% at the dosage of 0.80 microg/mL. Apoptosis rate caused by octreotide of non-transfected PC-3 cells was 5.76+/-0.75% at the dosage of 0.20 microg/mL, 6.69+/-0.80% at the dosage of 0.40 microg/mL and 7.26+/-1.28% at the dosage of 0.80 microg/mL. Transfected PC-3 cells growth inhibition rate caused by octreotide was 9.36+/-1.34% at the dosage of 0.20 microg/mL, 12.03+/-1.44% at the dosage of 0.40 microg/mL and 20.23+/-4.21% at the dosage of 0.80 microg/mL. Non-transfected PC-3 cells growth inhibition rate caused by octreotide was 6.44+/-0.66% at the dosage of 0.20 microg/mL, 7.65+/-0.88% at the dosage of 0.40 microg/mL and 9.29+/-1.32% at the dosage of 0.80 microg/mL. We found that octreotide caused higher apoptosis rate and inhibition rate in transfected groups than in non-transfected groups (P<0.05) at the tested dosages (0.20, 0.40 and 0.80 microg/mL). CONCLUSION: Deficiency of SST2 was probably the major reason why octreotide had little effect on PC-3 cells. Transfecting SST2 gene could strengthen the ability of octreotide of killing PC-3 cells. It provided an experimental evidence for using both octreotide and transfection with SST2 gene on clinical treatment of pancreatic cancer.

Effects of cyclooxygenase-2 antisense vector on proliferation of human cholangiocarcinoma cells.

OBJECTIVE: To transfect antisense vector of human cyclooxygenase-2 (COX-2) gene into COX-2 highly expressing cholangiocarcinoma cell line QBC939 and explore its biological activities and role in carcinogenesis. METHODS: QBC939 cells were transfected with antisense vector of human COX-2 gene using LipoVec transfecting technique. Transfected cells were selected with G418; COX-2 mRNA was examined using reverse transcription polymerase chain reaction (RT-PCR) and COX-2 protein expression was detected by immunocytochemistry using isozyme selective antibodies. The proliferative status of transfected cells was measured by using methabenzthiazuron (MTT) assay; Cell cycle and apoptosis were analyzed by using flow cytometry. RESULTS: RT-PCR showed a lower COX-2 mRNA level in antisense vector transfected cells and immunocytochemistry showed a weaker COX-2 protein expression in antisense vector transfected cells. The antisense vector transfected cells proliferative index decreased significantly (P < 0.01), the percentage of S phase decreased remarkably (P < 0.05) in antisense vector transfected cells (9.27% +/- 1.91%) compared with that in QBC939 cells without transfection(16.35% +/- 2.87%), and the percentage of G0/G1 phase increased remarkably (P < 0.05) in antisense vector transfected cells (75.16% +/- 4.13%) compared with that in QBC939 cells without transfection (57.31% +/- 10.16%). Transfection with antisense vector of human COX-2 gene had no significant influence on the apoptosis in QBC939 cells (P > 0.05). CONCLUSION: Transfection with antisense vector of human COX-2 gene could inhibit the proliferation of human cholangiocarcinoma QBC939 cells.