Human primary liver cancer-derived organoid cultures for disease modeling and drug screening.

Non-invasive liquid biopsies may transform the management of patients with cancer. Circulating tumour DNA (ctDNA) derived from cancer cells can be identified in most patients with advanced cancer, representing a potential non-invasive source of tumour DNA. The analysis of ctDNA may be useful to genotype the cancer, to select treatment options and to monitor response to treatment. ctDNA is often at a low level in plasma, requiring highly sensitive and accurate assays for ctDNA analysis. ctDNA was detected and profiled together with primary tumour DNA obtained from surgical specimens in 40 patients with lung cancer with localised disease who were diagnosed at stages I–III and operated on with curative intent. In a study carried out by investigators at the University of Stanford and recently published in Cancer Discovery,1 more than 90% of relapsing patients were found to have ctDNA 1 month after resection. This indicates a valuable detection of minimal residual disease. Moreover, ctDNA was able to show residual disease earlier than 5 months, compared with conventional follow-up imaging. The most frequently detected mutations in surveillance samples included mutations in TP53, KRAS, EGFR and KEAP1. Patients with detectable ctDNA at any post-treatment time point had significantly lower freedom from progression and survival than those in whom ctDNA was not detectable after completion of therapy. These findings suggest that ctDNA analysis is a promising approach for minimal residual disease detection in patients with localised lung cancers and that it can identify recurrence significantly earlier than routine CT imaging. The sensitivity and specificity of this diagnostic approach for detecting ctDNA were very high. In this series, all patients with detectable …

To examine the extent to which changes in diagnostic methods and classification are responsible for the striking increase in incidence of primary liver cancer in Denmark since 1943. Analysis of the time trends in sex specific, age standardised incidence of primary liver cancer and unspecified liver cancer (either secondary without known primary cancer or not specified as primary cancer) in the entire population from 1943 to 1985. By review of the 727 notifications from three periods of 5 years (1948-52, 1963-7, and 1978-82) the changes in histological diagnosis and classification were assessed. Denmark. Notifications of liver cancer to the Danish cancer registry. Concomitant with the increase in primary liver cancer, the incidence of the unspecified liver cancer declined. The proportion of histologically diagnosed primary liver cancer rose from 85% to 98%, whereas the proportion for unspecified liver cancer rose from 12% to 51%. When the proportion of primary versus unspecified liver cancer obtained by histological diagnosis was extrapolated to all cases, the annual incidence of primary liver cancer was 4.4 rather than 1.6 per 100,000 population in 1948-52 and 6.0 rather than 5.5 per 100,000 in 1978-82. The increase in the incidence of primary liver cancer may be much smaller than the numbers of registered cases indicate. This example emphasises the need to consider diagnostic drift in time trend studies of disease incidence.

Objective To explore the clinical diagnosis meaning and value of AFP on secondary liver cancer and primary liver cancer.Method Selected 87 patients diagnosed with liver cancer from July 2009to December 2010 in our hospital.Tested their serum AFP and recorded their serum AFP＞ 20 μ g/L,and ＞ 400μ g/L.When serum AEP ＞ 400 μ g/L,it is positive.Result 46 primary liver cancer patients' serum AFP＞20 μ g/L.39 patients were positive(serum AFP ＞400 μ g/L) ; positive rate was 81.25％.And among the secondary liver cancer patients,19 cases' serum alphafetoprotein ＞ 20 μ g/L; 10 cases' Serum AFP was positive; the positive rate was 25.64％.Primary liver cancer' serum alphafetoprotein positive rate was obviously higher than secondary liver cancer'.There was significant difference on positive diagnosis rate between the two groups(P＜0.05).Gonclusion Using serum AFP to diagnose primary liver cancer has a higher accuracy rate than to diagnose secondary liver cancer,but its sensitivity and specificity are insufficient ideal; assisting with other laboratory diagnosis can improve liver cancer diagnosis rate. Key words: Primary liver cancer (PHC); Secondary liver cancer(MHC); AFP clinical significance

The aim of this study was to explore the diagnostic value of liver stiffness measurement combined with serum high-sensitivity C-reactive protein detection in HBV-related cirrhosis patients complicated with primary liver cancer. A total of 156 previously untreated chronic hepatitis B-related cirrhosis patients and 50 healthy subjects were included in this study. The 156 patients were divided into two groups: those with primary liver cancer (67 cases) and without liver cancer (89 cases). The 50 healthy subjects were considered as normal control group. Liver stiffness measurement (LSM) was conducted and serum high-sensitivity C-reactive protein (CRP) level was assayed in all the 156 patients and 50 normal individuals, and their measurement values were statistically compared and analyzed. The LSM value was (39.72±29.05) kPa in the liver cancer patients, significantly higher than the (27.81±18.46) kPa in the cirrhosis alone patients and (4.25±0.74) kPa in the healthy controls (P<0.01 for both). Serum hs-CRP levels in the liver cancer patients was 5.81mg/L, significantly higher than 1.78 mg/L in the cirrhosis alone patients and 0.38mg/L in healthy controls, (P<0.01 for both). The higher the grade of LSM values was, the positive rate of CRP was higher in the cirrhosis patients complicated with primary liver cancer. In patients with LSM values ≥27.6 kPa, the serum CRP positive rate was 64.2% in patients with primary liver cancer, significantly higher than the 38.0% in patients with cirrhosis alone (P<0.01). In the 67 HBV-related cirrhosis patients complicated primary liver cancer, the LSM value and serum hs-CRP level in AFP-positive patients were (48.95±28.59) kPa and 4.91 mg/L, respectively, higher than those in the AFP-negative patients (28.64±26.83) kPa and 4.16 mg/L, but with a non-significant difference (P>0.05). Liver stiffness measurement combined with serum high-sensitivity C-reactive protein detection may have potential diagnostic implications as a marker of primary liver cancer occurrence in patients with HBV-related cirrhosis.

This retrospective analysis was conducted to evaluate the feasibility and safety of high-intensity focused ultrasound ablation for primary liver cancer and metastatic liver cancer. Patients with liver cancer who received high-intensity focused ultrasound were included in this analysis, including a primary liver cancer cohort (n=80) and a metastatic liver cancer cohort (n=195). The primary endpoint of our research was tumor response. The secondary endpoints included survival outcomes, visual analog scale pain scores, alpha-fetoprotein relief, and complications. Objective response rate and disease control rate were observed to be 71.8% and 81.2%, respectively, in patients with primary liver cancer and were 63.7% and 83.2% in cases with metastatic liver cancer. Alpha-fetoprotein levels and visual analogue scale levels significantly decreased after treatment compared with the baseline levels in patients with primary liver cancer (p<0.05). Median overall survival was estimated to be 13.0 and 12.0 months in the primary liver cancer and metastatic liver cancer cohorts. The 1-year survival rate was 70.69% and 48.00%, respectively. Multivariate regression analysis showed that visual analogue scale ≥ 5, longest diameter ≥ 5 cm, and portal vein invasion were the independent risk factors for poor survival in primary liver cancer. For patients with metastatic liver cancer, independent risk factors were identified as visual analogue scale ≥ 5, longest diameter ≥ 5 cm, existence of extrahepatic metastases, existence of portal vein invasion, and time to high-intensity focused ultrasound treatment from diagnosis < 3 months. Severe adverse events were rarely reported. In conclusion, high-intensity focused ultrasound might be an effective and safe option for patients with liver cancer regardless of primary and metastatic lesions.

Objective To investigate the significance of XAGE-1bmRNA expression in the blood specimens of patients with primary liver cancer, cirrhosis and benign liver diseases and in healthy individuals. Methods Venous blood specimens of patients with primary liver cancer (n= 125), cirrhosis (n= 23), benign liver diseases (n= 34) and healthy individuals (n = 41 ) were collected. XAGE-1b mRNA was detected by real-time fluorescence quantitative PCR. Results The expression levels of XAGE-1b mRNA in patients with primary liver cancer, cirrhosis, benign liver diseases and healthy in dividuals were 3.72 (0.93, 10.2) ×10-5, 0 (0, 0. 56) ×10-5, 0 (0, 0)×10-5, 0 (0, 0) ×10-5, respectively. The XAGE-1b mRNA expression in patients with primary liver cancer was obviously higher than the patients with cirrhosis, benign liver diseases and in healthy individuals. The expression levels for patients with cirrhosis was higher than patients with benign liver diseases and in healthy individuals. The expression levels for patients with benign liver diseases and healthy individuals were similar. With a optimal cut-off value of 8. 385 × 10-7 , the sensitivity, specificity, positive predictive value, and negative predictive value of XAGE-lb mRNA for diagnosing primary liver cancer were 80. 0%, 89.8%, 90.9% and 77.9% respectively. The positive rates for patients with primary liver cancer and cirrhosis were 80.0% and 30.4% respectively. Conclusion XAGE-lb mRNA can be used as a tumor marker for primary liver cancer. It contributes to the differentiation between primary liver cancer, cirrhosis and benign liver diseases. Key words: Liver carcinoma; Liver cirrhosis; Benign liver diseases; XAGE-1b mRNA

Background The drug-eluting beads transarterial chemoembolization (DEB-TACE) has already been used in hepatic malignancies. We aim to evaluate the efficacy and safety of DEB-TACE in treating primary or secondary liver cancer. Methods We retrospectively evaluated 59 patients with hepatic malignancies, including 41 patients with primary liver cancer and 18 patients with secondary liver cancer, between September 2016 and February 2019. All patients were treated with DEB-TACE. Objective response rate (ORR) and disease control rate (DCR) were evaluated by mRECIST. The pain was assessed using a numerical rating scale (NRS) where 0 represented no pain, and a score of ten was unbearable. Adverse reactions were assessed according to Common Terminology Criteria for Adverse Events 4.0 (CTCAE4.0). Results In the subgroup of primary liver cancer, 3 patients (7.32%) got complete response, 13 patients (31.71%) got partial response, 21 patients (51.22%) experienced stable disease, and 4 patients (9.76%) suffered progressive disease; ORR was 39.02% and DCR was 90.24%. In the subgroup of secondary liver cancer, 0 patients (0%) got complete response, 6 patients (33.33%) got partial response, 11 patients (61.11%) experienced stable disease, and 1 patient (5.56%) suffered progressive disease; ORR was 33.33% and DCR was 94.44%. We did not find any difference when comparing the efficacy between primary and secondary liver cancer (P=0.612). The one-year survival rate was 70.73% for primary liver cancer and 61.11% for secondary liver cancer. There was no significant difference between the two groups (P=0.52). For the patients with CR or PR, no factor could predict the efficacy of DEB-TACE. The most common treatment-related adverse reactions were short-term liver function disorders. The symptoms included fever (20.34%), abdomen pain (16.95%), and vomiting (5.08%), all patients with adverse reactions got remission after treatment. Conclusions DEB-TACE has a promising effect in the treatment of primary or secondary liver cancer. The treatment-related adverse reactions are tolerable.

Objective To summarize and analyze the surgical therapies for liver and gastric multiple primary cancers. Methods The clinical data of 14 patients with liver and gastric carcinomas surgically treated in our hospital from January 2004 to January 2008 were retrospectively analyzed.Results Of the 14 patients, 12 underwent simultaneous resection of liver and gastric carcinomas,1 resection of the gastric carcinoma 2 months after the liver surgery and 1 removal of liver cancer found 2 years after the surgery for antral adenocarcinoma. The median survival time of these patients was 23 months. The 1-and 3-year survival rate was 78.6% and 35.7%, respectively. Conclusion Due to different pathological characteristics, the therapies of liver and gastric multiple primary cancers are completely different from that of the recurring and metastatic carcinoma. Both tumors can be treated by radical resection and the effect is similar to single cancer. Positive treatment is crucial for long-term survival of the patients. Key words: Carcinoma, multiple primary; Primary Liver Cancer; Gastric Cancer; Surgery; Prognosis

Structure and expression of the cyclin A gene in human primary liver cancer. Correlation with flow cytometric parameters

The concentrations of N-terminal peptide of type III procollagen in the sera of patients with various cancers were measured by radioimmunoassay. The mean value (with standard deviation) in the control group was 9.9 +/- 2.6 ng/ml. Serum levels exceeding 15 ng/ml were defined as positive, and it was found that 94% of 18 patients with primary liver cancer with cirrhosis, 88% of 8 patients with primary liver cancer without cirrhosis, 77% of 13 patients with metastatic liver cancer, 86% of 7 patients with recurrent breast cancer, 86% of 8 patients with colonic cancer, 75% of 8 patients with pancreatic cancer, 70% of 23 patients with stomach cancer, 51% of 35 patients with lung cancer, and 54% of 28 patients with uterine cancer showed positive levels. The concentrations showed great intersubject variations, probably reflecting the activity of tumor growth and/or invasion. The concentrations in the sera of patients with primary liver cancer with cirrhosis were generally higher than those in patients with liver cirrhosis alone or primary liver cancer without cirrhosis. This result suggested that the growth of primary liver cancer complicated by cirrhosis might be detected by serial measurements of this peptide in the serum of patients with liver cirrhosis. Present data suggested that this peptide is not cancer-specific, but assay of the peptide might be of value as an auxiliary means of detecting and monitoring various cancers, especially liver cancer.

Objective To investigate the application value of three-dimensional surgery planning system in the preoperative evaluation of primary liver cancer (PLC) resection. Methods A total of 44 patients with PLC [32 males and 12 females, mean age of (60±12) years old] in Department of General Surgery, Lanzhou University Second Hospital from June 2012 to June 2013 were enrolled in this prospective study. The informed consents of all patients were obtained and the ethics committee approval was received. According to random number table method, the patients were randomly divided into 2 groups: computed tomography angiography (CTA) group and three-dimensional surgery planning (3D) group. Subgroups of complex PLC and non-complex PLC were further defined in each group according to tumor size, tumor invasive extent and history of surgery. In CTA group, 8 cases was assigned in complex PLC subgroup and 14 cases in non-complex PLC subgroup, and 6, 16 cases respectively in 3D group. CTA was used in the preoperative evaluation of PLC resection in CTA group. Liver three-dimensional surgery planning system was used in the preoperative evaluation of PLC resection in 3D group. The intra-operative finding was taken as a gold standard. The visualization of PLC, the adjacent relationship between PLC and peripheral tissues, the display rates of 12 abdominal vessels, variation of hepatic artery, vascular invasion of tumor, cholangiectasis by CTA and three-dimensional surgery planning system were observed. The relationship between estimated tumor volume by three-dimensional surgery planning system and actual weights of resected tumor was analyzed. Non-normal distribution data were expressed in M(Q25,Q75). The display rates by 2 methods were compared using Chi-square or Fisher's exact probability test. The relationship between estimated tumor volume by three-dimensional surgery planning system and actual weights of resected tumor was analyzed using Spearman rank correlation analysis. Results PLC lesions could be both visualized by 2 methods. The adjacent relationship between PLC and peripheral tissues could also be clearly visualized by three-dimensional surgery planning system. For patients with complex PLC, the display rate of abdominal vessels was 81% (78/96) in CTA group, and was 100% (72/72) in 3D group, where significant difference was observed (χ2=15.1, P<0.05). Cholangiectasis could not be visualized in CTA group and the display rate of cholangiectasis was 3/6 in 3D group. For patients with non-complex PLC, the display rate of abdominal vessels was 90% (151/168) in CTA group, and was 100% (192/192) in 3D group, where significant difference was observed (χ2=20.39, P<0.05). Cholangiectasis could not be visualized in CTA group and the display rate of cholangiectasis was 19% (3/16) in 3D group. For patients with complex PLC, the median estimated tumor volume by three-dimensional surgery planning system was 218(129,429)ml and the actual weights of resected tumor was 194(112,429)g, where positive correlation was observed (r =0.943, P<0.05) with an average error rate of 6.5%. For patients with non-complex PLC, the estimated tumor volume by three-dimensional surgery planning system was 368(89,560)ml and the actual weights of resected tumor was 395(126,578)g, where positive correlation was observed (r =0.958, P<0.05) with an average error rate of 6.3%. Conclusions Compared with CTA, three-dimensional surgery planning system can better display the adjacent relationship between tumor and peripheral tissues, abdominal vessels, cholangiectasis and estimate the volume of resected tumor more accurately. It is especially suitable for patients with complex liver cancer. Key words: Tomography, spiral computed; Angiography; Imaging, three-dimensional; Carcinoma, hepatocellular; Hepatectomy

Objective To detect the expression of extracellular matrix protein 1 ( ECM1 ) in primary liver cancer tissues, and explore its clinical significance in liver cancer metastasis. Methods Sixty cases of primary liver cancer tissues and adjacent tissues from 60 patients who were admitted to the Affiliated Provincial Hospital of Anhui Medical University from June 2008 to December 2009 were collected, and nine cases of normal liver tissues were collected from patients with liver trauma as control. The expression of ECM1 and the relationship between ECM1 and clinicopathological features of liver cancer were detected and analyzed using the immunohistochemistry and Western blot. All data were analyzed using the chi-square test, Fisher exact test and t test. Results ECM1 was mainly expressed in the cytoplasm of liver cells. The positive expression rate of ECM1 in liver cancer tissues was 73%, which was significantly higher than those in adjacent tissues (20%) and normal liver tissues (22%)( x2 = 34.286, 7. 044, P ＜ 0.05 ). The expression of ECM1 was correlated with liver cancer metastasis and TNM stages ( x2 = 5. 455, 4.275, P ＜ 0.05), while not with sex, age, size, capsule and differentiation of the tumor,alpha fetoprotein level and the expression of hepatitis B surface antigen ( x2 = 2. 841, 0. 014, 0. 000, 0. 734,0.075, 0.000, 0.031, P＞0.05). The result of Western blot indicated that the relative content of ECM1 in the liver cancer tissues was 25.49 ± 4.61, which was significantly higher than those in adjacent tissues (3.00 ±0.37) and normal liver tissues (2.94 ± 0.21 ) ( t = 31. 962, 31. 699, P ＜ 0. 05 ). Conclusions The expression of ECM1 in liver cancer tissues is higher than those in adjacent and normal liver tissues, and ECM1 expression is correlated with metastasis of liver cancer and TNM stages, which indicate that ECM1 may play a role in the metastasis of liver cancer, and it could be used as an indicator for liver cancer metastasis. Key words: Liver neoplasms; Extracellular matrix protein 1; Metastasis

Expression of insulin-like growth factor II (IGF-II) in human primary liver cancer: mRNA and protein analysis

Liver cancer is the fourth leading cause of cancer-related mortality and is distinguished by a relative paucity of chemotherapy options. It has been hypothesized that intratumor genetic heterogeneity may contribute to the high failure rate of chemotherapy. Here, we evaluated functional heterogeneity in a cohort of primary human liver cancer organoid lines. Each primary human liver cancer surgical specimen was used to generate multiple cancer organoid lines, obtained from distinct regions of the tumor. A total of 27 liver cancer lines were established and tested with 129 cancer drugs, generating 3,483 cell survival data points. We found a rich intratumor, functional (drug response) heterogeneity in our liver cancer patients. Furthermore, we established that the majority of drugs were either ineffective, or effective only in select organoid lines. In contrast, we found that a subset of drugs appeared pan-effective, displaying at least moderate activity in the majority of these cancer organoid lines. These drugs, which are FDA approved for indications other than liver cancers, deserve further consideration as either systemic or local therapeutics. Of note, molecular profiles, obtained for a reduced sample set, did not correlate with the drug response heterogeneity of liver cancer organoid lines. Taken together, these findings lay the foundation for in-depth studies of pan-effective drugs, as well as for functional personalized oncology approaches. Lastly, these functional studies demonstrate the utility of cancer organoid drug testing as part of a drug discovery pipeline.

Objective To investigate the clinicopathologic characteristics and prognosis of patients with lymph node metastasis of primary liver cancer. Methods The clinical data of 236 patients who underwent radical resection of primary liver cancer at the First Affiliated Hospital of Medical College of Xi'an Jiaotong University from August 2000 to August 2011 were retrospective analyzed. The clinicopathologic factors affecting survival time of patients with liver cancer, gender, age, pathological classification, histological grade, cirrhosis, alanine transami nase (ALT) , aspartate transaminase (AST) , alkaline phosphatase (ALP) , glutamyl-transpeptidase (GGT) , alpha-fetoprotein (AFP) , hepatitis, hepatic lobe distribution, tumor diameter, gross type, tumor number, satellite lesions, capsule integrity, liver capsule local invasion, vascular thrombosis, TNM stage and lymph node metastasis were analyzed. The patients received serum AFP and B ultrasound reexaminations every 3 months after operation and chest X-ray film every 6 months. The recurrence of liver cancer in suspected patients were confirmed by abdomen CT scan and/ or digital subtraction angiography (DSA) . The ultrasound-guided puncture histopatho-logical examination was done if necessary. The follow-up was done till 30, December 2013 or death. The survival time was calculated from operation date to death or the end of follow-up. The measurement data with normal distribution were presented as ±s and analyzed usingt test, and the count data were analyzed using chi-square test or Fisher exact probability. The survival curve was drawn by Kaplan Meier method, and the survival rate was analyzed using the Log rank test. The univariate analysis and multivariate analysis were done using the one-way ANOVA and COX regression model. Results Of 236 patients with primary liver cancer, there were 23 patients with lymph node metastasis and 213 patients without lymph node metastasis. The incidences of lymph node metastasis in patients with hepatocellular carcinoma, cholangiocarcinoma and hepatocellular cholangiocarcinoma were 7. 93%(13/ 164) , 15. 00% (9/ 60) and 8. 33% (1/ 12) , respectively. The lymph node metastasis which located in hepa-toduodenal ligament, hepatic pedicle, pancreatic and multiple locations were in 10, 6, 4 and 3 patients, respectively. Univariate analysis showed that the pathological classification, level of ALP, hepatitis, TNM stage, lymph node metastasis were risk factors affecting the prognosis of patients (F =3. 386, 4. 064, 2. 857, 22. 988, 4. 087, P 120 U/ L, TNM stage Ⅲ and positive lymph node metastasis were independent risk factors affecting the prognosis of patients (HR = 1. 533, 1. 592, 2. 032, 4. 086, 95% confidence interval: 1. 008-2. 331, 1. 019-2. 489, 1. 214-3. 399, 1. 996-8. 363, P 0.05) . Conclusions The hepatocellular carcinoma, ALP >120 U/ L, TNM stage Ⅲ and positive lymph node metastasis are independent risk factors affecting the prognosis of patients with primary liver cancer, and lymph node metastasis is closely related to liver lobe distribution of tumor, satellite lesions and partial liver capsule invasion. There is different in the prognosis of patients with positive lymph node metastasis and different pathological classification of primary liver cancer. Key words: Liver neoplasms; Pathology, clinical; Lymph node metastasis; Prognosis