The prognostic impact of microvascular invasion on patients with intrahepatic cholangiocarcinoma after R0 resections / 中华肝胆外科杂志

Objective To study the clinical impact of microvascular invasion (MVI) on patients with intrahepatic cholangiocarcinoma (ICC) after R0 resections.Methods The clinicopathological data of 359 patients with ICC who underwent R0 resection in the Zhongshan Hospital,Fudan University between January 2000 and December 2008 were retrospectively studied.Univariate analysis and multivariate analysis were carried out to study factors related to postoperative survival outcomes and recurrence.The impact of MVI on patients with ICC after R0 resection was studied.Results The incidence of MVI was 13.6％ in the study cohort.MVI was correlated with HBV infection (P ＜ 0.05),liver cirrhosis (P ＜ 0.05) and tumor differentiation (P ＜ 0.05).The 1-,3-,5-year overall survival (OS) between the MVI positive and negative groups were 50.0％,20.9％,12.2％ and 63.9％,33.1％,22.0％ respectively (P ＜ 0.05),and the median survival time was 13 months and 18.5 months (P ＜0.05).The 1-,3-,5-year recurrence free survival (RFS) rates between the MVI positive and negative groups were 29.7％,12.7％,8.5％ and 50.6％,26.9％,18.4％,respectively (P ＜0.05),and the median recurrence free survival time was 8 months and 12.5 months (P ＜ 0.05).Multivariate analysis showed that MVI was an independent risk factor affecting recurrence after R0 resection (HR 1.852,95％ CI:1.075 ～ 3.195,P ＜ 0.05).Conclusions The occurrence of MVI in ICC patients was associated with hepatitis B infection.MVI was an independent risk factor affecting recurrence in ICC patients after R0 resection.However,it was not an independent risk factor of overall survival in patients after R0 resection.The clinical impact of MVI on patients with ICC was not as strong as for hepatocellular carcinoma.

A nude mouse model of hepatocellular carcinoma single cell-derived organ site-specific metastasis / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To establish a single cell-derived organ site-specific metastatic model of human hepatocellular carcinoma (HCC) in the nude mouse.</p><p><b>METHODS</b>Using the limited dilution method, HCCLM3-R-LM1 and HCCLM3-R-LnM1 cell lines were used to generate eight (LM1-S2, -S3, -S4, -S5, -S11, -S15, -S21, and -S23) and five (LnM1-S7, -S11, -S13, -S17, and -S20) single cell-derived monoclonal cell lines, respectively. The monoclonal cell lines were seeded into 4-week-old nude mice, and three weeks later the resultant subcutaneous tumor tissues were orthotopically transplanted into the livers of nude mice. At six weeks after implantation, lung and lymph node were extracted for analysis of the metastatic foci fluorescence area and pathology to assess the number of metastatic foci.</p><p><b>RESULTS</b>Among the 13 mice implanted with the established monoclonal cell lines, six grew subcutaneous tumors. When orthotopically transplanted, the six tumors showed remarkably different metastatic potential and organ site-specific tropism. The fluorescence areas of lung metastatic foci were: LM1-S3, 80 923+/-10 162; LM1-S4, 1506 000+/-297 064; LM1-S5, 36 140+/-8 210; and LM1-S11, 508 448+/-134 272 (P less than 0.01); no lymph node metastases were found for these lines. For LnM1-S11, the fluorescence areas of lung and lymph node metastatic foci were 435 062+/-206 620 and 1 254 000+/-225 171, respectively.</p><p><b>CONCLUSION</b>We successfully established several monoclonal cell lines and nude mouse models of HCC with different metastatic potential and organ tropism. Among them, LM1-S3, LM1-S4, LM1-S5, and LM1-S11 have metastasis organotropism to lung. The LnM1-S11 line exhibits dual metastasis organotropism to lung and lymph node. These monoclonal cell lines and nude mouse models may represent useful tools for study of HCC metastasis organotropism.</p>

The establishment of a systematic site-specific metastasis model of human hepatocellular carcinoma in nude mouse / 中华肝脏病杂志

To establish a systematic site-specific metastatsis model of human hepatocellular carcinoma (HCC) in nude mouse. HCCLM3-R cells were seeded into mice liver to establish xenograft mouse models. With the help of RFP, metastasis foci in lungs and lymph nodes in mice were detected using fluorescent stereomicroscopy and were removed. Cells derived from the metastasis foci were named HCCLM3-R-LM1 and HCCLM3-R-LnM1 respectively. HCCLM3-R-LM1 and HCCLM3-R-LnM1 cells were seeded into mice livers to analyze the lung and lymph node metastasis. Lungs of all tested mice were collected, examined by pathological evaluation and counted lung metastasis. Both lung and lymph node metastasis were found in HCCLM3-R-LM1, HCCLM3-R and HCCLM3-R-LnM1 cells and a significant difference was found between the lung and the lymph node metastasis levels in the three cells. The fluorescent areas (pixels) of lung and lymph node metastasis were 8687.00+/-1844.63 versus 2570.00+/-318.20 (P = 0.0031) in HCCLM3-R-LM1 cells, 6457.67+/-832.62 versus 10 994.33+/-2 212.31 (P = 0.0036) in HCCLM3-R cells, and 2968.67+/-2571.00 versus 24 416.00+/-7 186.13 (P = 0.0094) in HCCLM3-R-LnM1 cells, respectively. The middle numbers of microscopic lung metastatic foci were 775, 430 and 310 in HCCLM3-R-LM1, HCCLM3-R and HCCLM3-R-LnM1 cells (P less than 0.001), respectively, consist with the results quantified by RFP. We established the systematic site-specific metastasis models which demonstrates lung- and lymph node-specific metastasis potential in nude mice and can be used as a model for researches on site-specific metastasis of HCC.

Effects of ARNT2 gene on invasion and migration of human hepatocellular carcinoma HCCLM6 cell line / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To investigate the effects of ARNT2 on invasion and migration of HCCLM6 cells.</p><p><b>METHODS</b>Four short hairpin oligos targeting to ARNT2 were s cloned into the pLVTHM vector. Lentiviral vectors shRNA-ARNT2i, pCMV-dR8.74 and pMD2G were cotransfected into 293T cells using Lipofectamine 2000. HCCLM6 was infected with virus supernatant. ARNT2 mRNA and protein expressions were detected using quantitative Real time-PCR and Western blot, respectively. The invasion and migration of HCCLM6 cells were evaluated using wound healing assay and cell invasion assay in vitro. Statistical analysis was performed with SPSS 16.0.</p><p><b>RESULTS</b>The relative mRNA levels of ARNT2 were 0.154+/-0.024, 0.860+/-0.145, 1.004+/-0.009 in shRNA-ARNT2i virus infected HCCLM6 cells, mock-infected cells and control vector virus infected cells (F = 113.14, P more than 0.01). The expression of ARNT2 at protein level was 16.45+/-1.6, 44.56+/-2.07 in the HCCLM6 cells infected with shRNA-ARNT2i virus and negative control vector virus, respectively (t = 18.58, P less than 0.01). The scrape wound of HCCLM6 cells infected with shRNA-ARNT2i virus healed faster than cells infected with control vector virus or mock-infected cells. The number of cells invading through Matrigel was higher in the HCCLM6 cells infected with shRNA-ARNT2i virus (13.25+/-1.04) than that in mock-infected HCCLM6 cells and the HCCLM6 cells infected with negative control vector virus (6.50+/-2.56, 6.75+/-2.05) (F = 29.645, P less than 0.01).</p><p><b>CONCLUSION</b>Inhibition of ARNT2 gene promotes the invasion and migration of HCCLM6 cells.</p>

Nuclear accumulation of CXCR4 and overexpressions of VEGF-C and CK19 are associated with a higher risk of lymph node metastasis in hepatocellular carcinoma / 中华肿瘤杂志

<p><b>OBJECTIVE</b>The aim of this study was to evaluate the correlation of protein expressions of CXC chemokine receptor 4 (CXCR4), vascular endothelial growth factor-C (VEGF-C) and cytokeratin 19 (CK-19) with lymph node metastasis (LNM) in patients with hepatocellular carcinoma (HCC), and their survival.</p><p><b>METHODS</b>The expressions of CXCR4, VEGF-C and CK-19 in HCC patients with (n = 123) or without (n = 145) LNM were determined using tissue microarray and immunohistochemical staining. The relationship between clinicopathological features and CXCR4, VEGF-C and CK-19 were analyzed. Evaluation of immunostaining was performed semiquantitatively by visual assessment.</p><p><b>RESULTS</b>The UICC T stage, and expressions of nuclear CXCR4, VEGF-C and CK-19 were independent risk factors for LNM. Nuclear CXCR4, VEGF-C and CK-19 expression were predictive factors for LNM in HCC patients. In patients with LNM, the median survival time was 15.1 months for patients with high nuclear CXCR4 expression and 24.5 months for those with low nuclear CXCR4 expression. The median survival time was 15.1 months for patients with high tumor VEGF-C expression and 31.1 months for those with low tumor VEGF-C expression. The median survival time was 12.0 months for patients with positive CK-19 expression and 19.2 months for patients with negative CK-19 expression. Patients with high nuclear CXCR4, VEGF-C or CK-19 expression had significantly poorer prognosis than those with low expression (all P < 0.05). PVT, UICC T stage and expressions of nuclear CXCR4, VEGF-C, and CK-19 were independent prognostic factors.</p><p><b>CONCLUSION</b>Increased protein expressions of nuclear CXCR4, VEGF-C, and CK-19 are independent risk factors for developing lymph node metastasis, and they are significantly correlated with LNM and poor outcome in HCC patients.</p>

A clinicopathologic study of hepatic angiomyolipoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the clinicopathologic features, diagnosis and treatment of hepatic angiomyolipoma (HAML).</p><p><b>METHODS</b>The clinical, histopathological, treatment and prognosis data of 51 patients treated for HAML from October 1998 to October 2007 were retrospectively analyzed.</p><p><b>RESULTS</b>HAML had a female predilection (female/male = 41/10) and the mean age was 44 years old. The main symptoms were abdominal mass (33 cases) and abdominal pain or discomfort (15 cases), the other 2 cases presented as fever. Histopathologically, HAML was composed of a heterogeneous mixture of blood vessels, smooth muscle, and adipose cells. Immunohistochemical staining showed relatively high positive rate of HMB-45 (50/51), SMA (47/49) and S-100 (39/42). All 51 patients underwent partial hepatectomy. The mean hospital stay was 13.8 days and mean intraoperative blood loss was 263 ml. There was no recurrence or metastasis after a mean follow-up of 55.4 months.</p><p><b>CONCLUSIONS</b>HAML is a rare benign mesenchymal tumor of the liver. Definitive diagnosis of HAML depends on the pathohistological findings and HMB-45 positive myoid cell is an important diagnostic marker. Complete surgical resection is the optimal treatment for HAML with favorable prognosis.</p>

Effects of high metastatic potential hepatocellular carcinoma cell-binding peptide on the invasion and metastasis of liver cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the effects of specific peptide (AWYPLPP peptide) binding to high metastatic potential human hepatocellular carcinoma (HCC) cells on the invasion and metastasis of liver cancer.</p><p><b>METHODS</b>The effects of AWYPLPP peptide on the invasion, migration, proliferation and adhesion of high metastatic potential human HCC cell line (HCCLM3) were evaluated in vitro by Matrigel invasion assay, migration assay, MTT assay and adhesion assay. The effect of AWYPLPP peptide on lung metastasis of HCC in vivo was evaluated in male nude mice with subcutaneously implanted HCCLM3 cells.</p><p><b>RESULTS</b>Incubation with the AWYPLPP peptide, but not the control peptide, resulted in a concentration-dependent increase of invasion ability in HCCLM3 cells at the concentration of 0.1 to 100 micromol/L. At any concentration used for the invasion assay, the peptide had no effect on cell migration, proliferation and adhesion. After 30 days of transplantation, eight of nine (88.9%) mice in the AWYPLPP peptide group showed obvious lung metastasis. The metastatic rate of lung metastasis was significantly increased in the AWYPLPP peptide group compared with that in the control group. There was no significant difference among the weights of primary tumor in the PBS, control peptide and AWYPLPP peptide groups.</p><p><b>CONCLUSION</b>AWYPLPP peptide can promote in vitro invasion and in vivo lung metastasis of high metastatic potential human HCC cells. Identification of the receptor for AWYPLPP peptide binding may provide new insights into the molecular mechanism underlying HCC invasion and metastasis as well as new targets for intervention.</p>

Primary experience of the anatomical laparoscopic left lateral hepatic lobectomy procedure for benign and malignant liver tumors / 中华外科杂志

<p><b>OBJECTIVE</b>To assess the feasibility, safety and outcome of anatomical laparoscopic left lateral hepatic lobectomy for benign and malignant liver tumors.</p><p><b>METHODS</b>From April 2005 to May 2008, 11 patients (7 male, 4 female; mean age 51.7 years) underwent anatomical laparoscopic left lateral hepatic lobectomy. Four patients presented with hepatocellular carcinoma and cirrhosis, while 1 patient had metastatic liver tumors from postoperatively colon cancer, five patients had hemangioma (2 cases with gallstones underwent cholecystectomy), 1 patient had a huge symptomatic angiolipoleiomyoma. Mean tumor size was 5.8 cm (range 2.1 to 12.0 cm). All the lesions were localized in the anatomical left lateral lobe (segments II to III).</p><p><b>RESULTS</b>The mean operative time was 147 min (range 120 to 180 min). There were no intraoperative or postoperative complications, and blood transfusions were not required. The mean postoperative hospital stay was 5.9 days.</p><p><b>CONCLUSIONS</b>Anatomical laparoscopic left lateral hepatic lobectomy are feasible and safety.</p>

Diagnosis and treatment of primary hepatic carcinoid tumor / 中华外科杂志

<p><b>OBJECTIVE</b>To discuss the diagnosis and treatment of primary hepatic carcinoid tumor (PHCT).</p><p><b>METHODS</b>Report one case of huge PHCT treated in February 2004, and search the other 19 cases which were published from January 1994 to December 2006 in the Chinese biological and medical literature database. The clinical manifestation, pathological findings, diagnosis and treatment of these 20 PHCT patients were analyzed retrospectively.</p><p><b>RESULTS</b>The main symptoms were abdominal pain or discomfort (8 cases) and abdominal mass (7 cases), cases with typical carcinoid syndrome were rare (3 cases). Immunohistochemical staining was positive for neuron-specific enolase, chromogranin A and synaptophysin in most cases. Sixteen cases received operation, among which there were 13 removed completely, other 4 cases were treated by transcatheter arterial chemoembolization (TACE).</p><p><b>CONCLUSIONS</b>The definite diagnosis of PHCT depends on pathological and histochemical findings. Complete surgical resection is the best treatment for PHCT with favourable prognosis. TACE is also effective for nonoperative cases.</p>

Transcription factor activity profile of human hepatocellular carcinoma cell lines with different metastatic potentials / 中华肝脏病杂志

<p><b>OBJECTIVES</b>To examine the activities of transcription factors (TFs) in human hepatocellular carcinoma (HCC) cell lines with different metastatic potentials, so as to identify the TFs associated with HCC metastasis.</p><p><b>METHODS</b>Transcription factor activity profile of Hep3B, MHCC97L and MHCC97H, three HCC cell lines with different metastatic potentials, were examined using protein/DNA array. Electrophoretic mobility shift assays (EMSA) and Western blot were used to confirm the results obtained by protein/DNA array.</p><p><b>RESULTS</b>From a total of 345 screened TFs, 7 activity differential TFs were found, of which 5 showed increased activity, including p53, hypoxia inducible factor-1 alpha (HIF-1alpha), signal transducer and activator of transcription 3 (Stat3) and Sp1, and 2 showed decreased activity including Rb and Smad3.</p><p><b>CONCLUSION</b>The abnormal functioning of transcription factors is closely associated with HCC metastasis. Our present findings could be of help in expanding our understanding of the mechanism of HCC metastasis and identify new predictive biomarkers and therapeutic targets.</p>

Reconstructing the JAK/STATs signal pathway restored the anti-proliferative response of MHCC97 on interferon alpha / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To elucidate the roles of JAK/STATs signal pathway on anti-proliferative effects induced by IFN-alpha in MHCC97.</p><p><b>METHODS</b>An IRF9 expression vector was transfected into MHCC97 with Dosper. The expression of IRF9, cycle regulating proteins and the forming of ISGF3 complex were detected using Western blot and EMSA, respectively. Cell proliferation and distribution were monitored using MTT and flow cytometry.</p><p><b>RESULTS</b>High expression of IRF9 restored the anti-proliferative response of MHCC97 on IFN-alpha treatment and delayed the cell transition from S phase to G2 phase induced by IFN-alpha.</p><p><b>CONCLUSION</b>The integrity and functions of JAK/STATs signal pathway played an important role in mediating the anti-proliferative effects of IFN-alpha in MHCC97.</p>

The prognostic significance of preoperative plasma level of osteopontin in combination with intercellular adhesion molecule-1 for patients with hepatocellular carcinoma / 中华外科杂志

<p><b>OBJECTIVE</b>To investigate the enhanced predictive activity of preoperative plasma osteopontin (OPN) level in combination with intercellular adhesion molecule-1 (ICAM-1) for recurrence and prognosis of patients after resection of hepatocellular carcinoma (HCC).</p><p><b>METHODS</b>A total of 75 patients received liver resection for HCC from August 2001 to December 2001 in authors' institute were enrolled in this study. The preoperative plasma levels of OPN and ICAM-1 were detected by ELISA, and the association of them combination with the recurrence and prognosis of HCC patients was analyzed.</p><p><b>RESULTS</b>OPN and ICAM-1 could be detected in all of the plasma samples of the tested patients. A significantly higher OPN level and ICAM-1 level were found in plasma of patients who were found to have HCC recurrence during the follow-up time compared with those without recurrence (210.40 vs. 154.86 ng/ml, P = 0.001; 1011.23 vs. 747.49 ng/ml, P = 0.027). A significant difference of OS and DFS were found in different subgroups with higher or lower level of OPN (625 vs. 808 days, P = 0.0006; 433 vs. 674 days, P = 0.0003); and a similar situation was found in patients of high- and low- ICAM-1 levels (651 vs. 794 days, P = 0.0269; 489 vs. 642 days, P = 0.0248). The 2-year recurrence rates of the patients with higher and lower plasma levels of both OPN and ICAM-1 were 87.50% and 28.00% (P < 0.001), respectively; and the 2-year OS rates were 37.50% and 88.00% (P = 0.001), and the 2-year DFS rates were 12.50%, and 76.00 (P = 0.001), respectively.</p><p><b>CONCLUSIONS</b>The evaluation of preoperative plasma level of OPN or ICAM-1 may be helpful to predict the recurrence and prognosis of HCC patients in advance. The assessment of OPN level in combination with ICAM-1 could stratify patients into groups with different potentials of HCC recurrence and different outcomes more accurately than OPN or ICAM-1 individually.</p>

Effect of postoperative transcatheter arterial chemoembolization on hepatocellular carcinoma patients with residual tumor / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To evaluate the effect of postoperative adjuvant transcatheter arterial chemoembolization (TACE) on hepatocellular carcinoma (HCC) patients with residual tumor.</p><p><b>METHODS</b>The patients were classified into intervention group (with adjuvant TACE) and control group (without adjuvant TACE) who were further stratified to those with high risk (patients with single tumor > 5 cm in diameter, or with multiple tumors, invasion to blood vessels), and low risk factors. Univariate analysis and Cox model were used to analyse prognostic factors.</p><p><b>RESULTS</b>In low risk patients with residual tumor, the 1-, 2-, 3-, 4-year survival rate was 97.2%, 78.0%, 66.5% and 66.5% in the intervention group, and 91.2%, 81.4%, 70.3% and 54.4% in the control group, respectively. There was no statistical difference between the two groups in survival (log-rank P = 0.7667). Comparing with the control group, the 1-, 2-, 3-, 4-year survival rate was 89.5%, 73.4%, 59.2% and 53.8% in the intervention group, and 70.5%, 61.9%, 46.8% and 46.8% in the control group, respectively. Postoperative adjuvant TACE significantly prolonged the survival in high risk patients with residual tumor (P = 0.0029). Cox model revealed that the benefit of adjuvant TACE was significantly increased by the high risk factors in HCC patients with residual tumor.</p><p><b>CONCLUSION</b>The beneficial effect of postoperative TACE was only observed in high risk patients with residual tumor but not in the low risk patients with residual tumor.</p>

Three-grade criteria of curative resection for primary liver cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To clarify three-grade criteria of curative resection for primary liver cancer (PLC) and evaluate their clinical significance.</p><p><b>METHODS</b>Criteria of curative resection of PLC were summed up to three grades. Grade I: complete removal of all gross tumors with no residual tumor at the excision margin. Grade II: on the basis of Grade I, there was no extrahepatic metastasis, no hilar lymph node metastasis, no tumor thrombus in the main trunks and their primary tributaries of the portal vein, common hepatic duct, hepatic vein and vena cava inferior, and the tumor was not more than two in number. Grade III: in addition to the above criteria, AFP dropped to normal level (in patients with elevated AFP before surgery) within 2 months after operation, and no residual tumor upon diagnostic imaging. A total of 354 cases with PLC who had their liver resected was reviewed. Patients in each grade were divided into two portions depending on whether the treatment was curative or palliative.</p><p><b>RESULTS</b>The survival of patients receiving curative treatment was better than those receiving palliative treatment (P < 0.01). This was true for patients whose treatment belonged to anyone of the three-grade criteria. The survival was improved along with the promotion of curative criteria used. The 5-year survival rate of Grade I, II and III patients undergone curative resection was 43.2%, 51.2% and 64.4%, respectively (P < 0.01).</p><p><b>CONCLUSION</b>1. The three-grade criteria may be used for judging the radicality of tumor resection for PLC. 2. The more stringent the criteria used, the better the survival would be. 3. Adopting high-grade criteria to select cases, to guide operation and postoperative follow-up would improve the results of liver resection for PLC.</p>