Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like m...Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like metastatic model of human HCC in nude mice (LCI-D20) and a low metastatic model of human HCC in nude mice (LCI-D35) have been established. All mice with transplanted LCI-D20 tumors exhibited extremely high metastatic ability including spontaneous metastasis to liver, lungs, lymph nodes and peritoneal seeding. Remarkable difference was also found in expression of some of the invasiveness related genes and growth factors between the LCI-D20 and LCI-D35 tumors. PAI-1 increased gradually following tumor progression in LCI-D20 model, and correlated with tumor size and AFP level. Phasic expression of tissue intercellular adhesion molecule-1 in this model was also observed. Using corneal micropocket model, it was demonstrated that the vascular response induced by LCI-D20 tumor was stronger than that induced by LCI-D35 tumor. Similar report on metastatic human HCC model in nude mice and human HCC cell line with metastatic potential was rarely found in the literature. This LCI-D20 model has been widely used for the studies on intervention of metastasis, including anti-angiogenesis,antisense approach, metalloproteinase inhibitor, differentiation inducer, etc. It is concluded that the establishment of metastatic human HCC model in nude mice and human HCC cell line with metastatic potential will provide important models for the in vitro and in vitro study of HCC invasiveness, angiogenesis as well as intervention of HCC recurrence.展开更多
Objective: To study relationship between apoptosis and invasive and metastatic potential of hepatocellu- lar carcinoma (HCC). Methods: Apoptotic rate (AR), proliferative index (PI) and S-phase fraction (SPF) were meas...Objective: To study relationship between apoptosis and invasive and metastatic potential of hepatocellu- lar carcinoma (HCC). Methods: Apoptotic rate (AR), proliferative index (PI) and S-phase fraction (SPF) were measured by flow cytometry, and p170, p21 and nucleoside diphosphate kinase (ndpk) by strept avidin-biotin complex immunohistochemical technique in 57 pa- tients with HCC. Results: In this group, AR was 1.77% ±0.19%, SPF 12.55% ±0.68%, and PI 20.91% ±1.12% (r =-0.173). p170, p21 and ndpk positive rates were 61.36%, 68, 18%, 52.27% respectively in patients with a mean AR of ≤1.77%, and 23.08%, 38.46%, 84.62% respectively in patients with a mean AR of >1.77% (all P<0.05). In patients with positive tumor invasiveness and metastasis, nd- pk (+) was 43.75%, p21 (+) 75.00%, p170 (+) 65.63%, AR 1.12% ±0. 16%, PI 23.78% ±1.48%, and SPF 13.90 % ±0.99 %. In patients with negative invasiveness and metastasis, however, ndpk (+) was 80.00%, p21 (+) 44.00%, p170 (+) 36.00%, AR 2,32%±0.52%, PI 18.53% ±0.82% and SPF 11.43% ±0.70%. Conclusion: Apoptosis of HCC is negatively correla- ted with its invasive and metastatic potential or other factors as proliferative activity, p21, p170 and ndpk.展开更多
Background： Biotherapy based on human bone marrow-derived mesenchymal stem cells （BMSCs） is currently the focus of research, especially in the field of autologous stem cell transplantation. A novel type of metastas...Background： Biotherapy based on human bone marrow-derived mesenchymal stem cells （BMSCs） is currently the focus of research, especially in the field of autologous stem cell transplantation. A novel type of metastasis-associated magnetic resonance （MR） molecular imaging probe was constructed, and the changes in metastasis and proliferation of hepatocellular carcinoma （HCC） before and after BMSC intervention were observed through MR imaging （MRI）. Methods：Metastasis-associatedMRmolecularimagingprobe,integrin αvβ3 ligandcRGD-PEG-DGL-DTPA-Gd （Gd-RGD）,wereconstructed. After human BMSC intervention was performed for 6weeks, tumor weight inhibition rates were calculated, and the RGD molecular probe was imaged through MRI with molecular imaging agent Gd-DTPAas control.The signal-to-noise ratio （SNR） and contrast-to-noise ratio （CNR） in the MRI experiment were used as semi-quantitative indicators. Polymerase chain reaction method was performed to detect proliferation- and metastasis-associated indicators, transforming growth factor β-1 （TGFβ1）, osteopontin （OPN）, and integrin subunit αv and β3. Results： The highest tumor weight inhibition rates were observed 3 weeks after the BMSC transplantation. The MR Gd-RGD in the HCC tissues after the BMSC intervention showed less enhancement than Gd-DTPA. The Gd-DTPAMRI of control group had higher SNR and CNR than Gd-RGD MRI in the experimental groups （P 〈 0.05）. For high metastatic potential hepatocellular carcinoma （MHCC97-H）, significant differenceswereobservedintheSNRsandCNRsofGd-RGDMRIbeforeandaftertheBMSCintervention（P〈0.05）.Forlowmetastaticpotential hepatocellular carcinoma （MHCC97-L）, the CNRs of Gd-RGD MRI were statistically different before and after BMSC intervention （P 〈 0.05）. With regard to MHCC97-H, OPN, β3, and TGFβ1 expression significantly decreased after BMSC intervention （P 〈 0.05）. In MHCC97-L and OPN, β3, TGFβ1, and αv expression after BMSC intervention decreased, and the difference was statistically significant （P 〈 0.05）. Conclusions： The CNR index of MRI is a good indicator for distinguishing high- and low-metastatic potential HCC tissues.After BMSC transplantation of MRI through the two kinds of tracer, the SNR and CNR indexes can distinguish two kinds of high and low metastatic potential HCC tissues, and Gd-RGD imaging is more suitable in distinguishing the metastatic potential changes through BMSC intervention.展开更多
In order to explore the genomic basis for liver cancer metastasis,whole-exome sequencing(WES)was performed on patient-derived hepatocellular carcinoma(HCC)cell lines with differential metastatic potentials and analyze...In order to explore the genomic basis for liver cancer metastasis,whole-exome sequencing(WES)was performed on patient-derived hepatocellular carcinoma(HCC)cell lines with differential metastatic potentials and analyzed their clonal evolution relationships.An evolutionary tree based on genomic single nucleotide polymorphism(SNP)was constructed in MegaX software.The WES data showed that the average percentage of heterogeneous mutations in each HCC cell lines was 16.55%(range,15.38%e18.17%).C:G>T:A and T:A>C:G somatic transitions were the two most frequent substitutions.In these metastatic HCC cell lines,non-silent gene mutations were found in 21.88%of known driver genes and 10 classical signaling pathways.The protein interaction network was constructed by STRING,and hub genes were found in the shared trunk mutation genes and the heterogeneous branch mutations respectively.In cBioPortal database,some of the selected hub genes were found to be associated with poor overall survival(OS)of HCC patients.Among the mutated HCC driver genes,a novel KEAP1 mutation with a homozygous frameshift truncation at the c-terminal Nrf2 binding region was detected and verified in MHCC97-H and HCC97LM3 cells.In conclusion,WES data demonstrate that HCC cell lines from tumor biopsy specimens of the same patient have obtained different metastatic potentials through repeated selection in rodents in vivo,and they do indeed have a genetic relationship at the genomic level.展开更多
基金Partly supporled by the State Key Basic Research Program Grant of China(G1998051211)Leading Speciality Grant of Shanghai Health Bureau.
文摘Metastatic human HCC model is needed for the studies on mechanism and intervention of metastatic recurrence. By using orthotopic implantation of histologically intact tissues of 30 surgical specimens, a patient-like metastatic model of human HCC in nude mice (LCI-D20) and a low metastatic model of human HCC in nude mice (LCI-D35) have been established. All mice with transplanted LCI-D20 tumors exhibited extremely high metastatic ability including spontaneous metastasis to liver, lungs, lymph nodes and peritoneal seeding. Remarkable difference was also found in expression of some of the invasiveness related genes and growth factors between the LCI-D20 and LCI-D35 tumors. PAI-1 increased gradually following tumor progression in LCI-D20 model, and correlated with tumor size and AFP level. Phasic expression of tissue intercellular adhesion molecule-1 in this model was also observed. Using corneal micropocket model, it was demonstrated that the vascular response induced by LCI-D20 tumor was stronger than that induced by LCI-D35 tumor. Similar report on metastatic human HCC model in nude mice and human HCC cell line with metastatic potential was rarely found in the literature. This LCI-D20 model has been widely used for the studies on intervention of metastasis, including anti-angiogenesis,antisense approach, metalloproteinase inhibitor, differentiation inducer, etc. It is concluded that the establishment of metastatic human HCC model in nude mice and human HCC cell line with metastatic potential will provide important models for the in vitro and in vitro study of HCC invasiveness, angiogenesis as well as intervention of HCC recurrence.
基金This project was financially supported by "9.5" National Research Project of China (No. 969070301) and the National Natural Science Foundation of China (No. 30070235).
文摘Objective: To study relationship between apoptosis and invasive and metastatic potential of hepatocellu- lar carcinoma (HCC). Methods: Apoptotic rate (AR), proliferative index (PI) and S-phase fraction (SPF) were measured by flow cytometry, and p170, p21 and nucleoside diphosphate kinase (ndpk) by strept avidin-biotin complex immunohistochemical technique in 57 pa- tients with HCC. Results: In this group, AR was 1.77% ±0.19%, SPF 12.55% ±0.68%, and PI 20.91% ±1.12% (r =-0.173). p170, p21 and ndpk positive rates were 61.36%, 68, 18%, 52.27% respectively in patients with a mean AR of ≤1.77%, and 23.08%, 38.46%, 84.62% respectively in patients with a mean AR of >1.77% (all P<0.05). In patients with positive tumor invasiveness and metastasis, nd- pk (+) was 43.75%, p21 (+) 75.00%, p170 (+) 65.63%, AR 1.12% ±0. 16%, PI 23.78% ±1.48%, and SPF 13.90 % ±0.99 %. In patients with negative invasiveness and metastasis, however, ndpk (+) was 80.00%, p21 (+) 44.00%, p170 (+) 36.00%, AR 2,32%±0.52%, PI 18.53% ±0.82% and SPF 11.43% ±0.70%. Conclusion: Apoptosis of HCC is negatively correla- ted with its invasive and metastatic potential or other factors as proliferative activity, p21, p170 and ndpk.
文摘Background： Biotherapy based on human bone marrow-derived mesenchymal stem cells （BMSCs） is currently the focus of research, especially in the field of autologous stem cell transplantation. A novel type of metastasis-associated magnetic resonance （MR） molecular imaging probe was constructed, and the changes in metastasis and proliferation of hepatocellular carcinoma （HCC） before and after BMSC intervention were observed through MR imaging （MRI）. Methods：Metastasis-associatedMRmolecularimagingprobe,integrin αvβ3 ligandcRGD-PEG-DGL-DTPA-Gd （Gd-RGD）,wereconstructed. After human BMSC intervention was performed for 6weeks, tumor weight inhibition rates were calculated, and the RGD molecular probe was imaged through MRI with molecular imaging agent Gd-DTPAas control.The signal-to-noise ratio （SNR） and contrast-to-noise ratio （CNR） in the MRI experiment were used as semi-quantitative indicators. Polymerase chain reaction method was performed to detect proliferation- and metastasis-associated indicators, transforming growth factor β-1 （TGFβ1）, osteopontin （OPN）, and integrin subunit αv and β3. Results： The highest tumor weight inhibition rates were observed 3 weeks after the BMSC transplantation. The MR Gd-RGD in the HCC tissues after the BMSC intervention showed less enhancement than Gd-DTPA. The Gd-DTPAMRI of control group had higher SNR and CNR than Gd-RGD MRI in the experimental groups （P 〈 0.05）. For high metastatic potential hepatocellular carcinoma （MHCC97-H）, significant differenceswereobservedintheSNRsandCNRsofGd-RGDMRIbeforeandaftertheBMSCintervention（P〈0.05）.Forlowmetastaticpotential hepatocellular carcinoma （MHCC97-L）, the CNRs of Gd-RGD MRI were statistically different before and after BMSC intervention （P 〈 0.05）. With regard to MHCC97-H, OPN, β3, and TGFβ1 expression significantly decreased after BMSC intervention （P 〈 0.05）. In MHCC97-L and OPN, β3, TGFβ1, and αv expression after BMSC intervention decreased, and the difference was statistically significant （P 〈 0.05）. Conclusions： The CNR index of MRI is a good indicator for distinguishing high- and low-metastatic potential HCC tissues.After BMSC transplantation of MRI through the two kinds of tracer, the SNR and CNR indexes can distinguish two kinds of high and low metastatic potential HCC tissues, and Gd-RGD imaging is more suitable in distinguishing the metastatic potential changes through BMSC intervention.
基金This work was supported by the National Natural Science Foundation of China(NSFC,NO.81172066,NO.81472858NO.91529103)+1 种基金Innovation Team Fund of Second Affiliated Hospital of Chongqing Medical UniversityThe authors would like to thank Dr.Zhou-You Tang,Professor&Director,Liver Cancer Institute,Fudan University,for providing the three HCC cell lines(MHCC97-L,MHCC97-H,HCC97LM3).
文摘In order to explore the genomic basis for liver cancer metastasis,whole-exome sequencing(WES)was performed on patient-derived hepatocellular carcinoma(HCC)cell lines with differential metastatic potentials and analyzed their clonal evolution relationships.An evolutionary tree based on genomic single nucleotide polymorphism(SNP)was constructed in MegaX software.The WES data showed that the average percentage of heterogeneous mutations in each HCC cell lines was 16.55%(range,15.38%e18.17%).C:G>T:A and T:A>C:G somatic transitions were the two most frequent substitutions.In these metastatic HCC cell lines,non-silent gene mutations were found in 21.88%of known driver genes and 10 classical signaling pathways.The protein interaction network was constructed by STRING,and hub genes were found in the shared trunk mutation genes and the heterogeneous branch mutations respectively.In cBioPortal database,some of the selected hub genes were found to be associated with poor overall survival(OS)of HCC patients.Among the mutated HCC driver genes,a novel KEAP1 mutation with a homozygous frameshift truncation at the c-terminal Nrf2 binding region was detected and verified in MHCC97-H and HCC97LM3 cells.In conclusion,WES data demonstrate that HCC cell lines from tumor biopsy specimens of the same patient have obtained different metastatic potentials through repeated selection in rodents in vivo,and they do indeed have a genetic relationship at the genomic level.
