BACH1 promotes the progression of human colorectal cancer through BACH1/CXCR4 pathway.

The present study was to investigate clinical significance, biological functions and underlying mechanisms of BTB Domain and CNC Homolog 1(BACH1) deregulation in human colorectal cancer (CRC). The result showed that BACH1 upregulation was significantly associated with enhanced tumor invasion (P = 0.014) and gender (P = 0.028) of CRC patients. Kaplan-Meier method results showed that the overall survival of CRC patients with high BACH1 mRNA expression was markedly shorter than those with low expression (P = 0.015), and multivariate analyzes results showed that BACH1 was an independent prognostic predictor for CRC patients (P = 0.049). In vitro studies revealed that BACH1 efficiently promoted invasion and migration of CRC cell line. In vitro studies proved that the HCT116 cell stably expressing BACH1 formed significantly larger tumor nodules and remarkably accelerated tumor xenografts growth. In addition, Immunohistochemical scores of CD31 and Vimentin were significantly higher than those of the control group. Finally, correlation analysis indicated that BACH1 expression was positively correlated with C-X-C Motif Chemokine Receptor 4(CXCR4) in tumor tissues and cell lines. Together, BACH1 serves as an oncogene to promote CRC progression and an independent prognostic factor for survival and metastasis. BACH1 may inhibit the progression of CRC through BACH1/CXCR4 pathway.

m6A-modified circFNDC3B inhibits colorectal cancer stemness and metastasis via RNF41-dependent ASB6 degradation.

Colorectal cancer (CRC) is the third most frequently diagnosed cancer with unfavorable clinical outcomes worldwide. circFNDC3B plays as a tumor suppressor in CRC, however, the mechanism of circFNDC3B in CRC remains ambiguous. The stem-like properties of CRC cells were detected by the evaluation of stemness markers, sphere formation assay and flow cytometry. qRT-PCR, FISH, IHC, and western blotting assessed the expression and localization of circFNDC3B, RNF41, ASB6, and stemness markers in CRC. The metastatic capabilities of CRC cells were examined by wound healing and Transwell assays, as well as in vivo liver metastasis model. Bioinformatics analysis, RNA immunoprecipitation (RIP), RNA pull-down assay and co-IP were used to detect the associations among circFNDC3B, FXR2, RNF41, and ASB6. Downregulated circFNDC3B was associated with unfavorite survival in CRC patients, and circFNDC3B overexpression suppressed CRC stemness and metastasis. Mechanistically, studies revealed that YTHDC1 facilitated cytoplasmic translocation of m6A-modified circFNDC3B, and circFNDC3B enhanced RNF41 mRNA stability and expression via binding to FXR2. circFNDC3B promoted ASB6 degradation through RNF41-mediated ubiquitination. Functional studies showed that silencing of RNF41 counteracted circFNDC3B-suppressed CRC stemness and metastasis, and ASB6 overexpression reversed circFNDC3B- or RNF41-mediated regulation of CRC stemness and metastasis. Elevated ASB6 was positively correlated with unfavorite survival in CRC patients. In vivo experiments further showed that circFNDC3B or RNF41 overexpression repressed tumor growth, stemness and liver metastasis via modulating ASB6. Taken together, m6A-modified circFNDC3B inhibited CRC stemness and metastasis via RNF41-dependent ASB6 degradation. These findings provide novel insights and important clues for targeted therapeutic strategies of CRC.

Normalization of T cell receptor repertoire diversity in patients with advanced colorectal cancer who responded to chemotherapy.

To investigate the correlation between normalization of T cell receptor (TCR) repertoire and remission of advanced colorectal cancer. Forty-one patients were randomly assigned to receive either folinic acid/fluorouracil/irinotecan alone (n = 20) or folinic acid/fluorouracil/irinotecan in combination with recombinant human endostatin (n = 21). Efficacy and toxicity were evaluated, and changes in TCR repertoire diversity were assessed by detecting the spectratypes of TCR complementarity-determining region three before and after several cycles of therapy. A scoring system was used to quantify changes in the TCR repertoire over time. The results demonstrated that the TCR repertoire exhibited a higher degree of normalization among patients undergoing remission relative to patients experiencing tumor progression. The results of the current study showed a positive correlation between TCR repertoire normalization and remission of colorectal cancer, suggesting that dynamic monitoring of TCR repertoire diversity may have potential prognostic value in the clinical setting.

Cancer of the gastrointestinal tract results in a restricted T-cell repertoire dependent upon tumor differentiation.

We investigated the influence of tumor tissue differentiation on the diversity of TCR repertoire. CDR3 spectratypes of CD4(+) and CD8(+) T cell subsets were analyzed from 27 patients with gastrointestinal tract tumors exhibiting varying degrees of differentiation. A CDR3 spectratype complexity scoring system was used to quantify the diversity of TCR repertoire. Each patient was matched with an age-matched healthy group to control for age variability. Results show that the complexity scores (TCR repertoire diversity) have a significant correlation with the degree of tumor differentiation, which provides useful information for understanding immune response in cancer patients.

Dynamic monitoring the TCR CDR3 spectratypes in patients with metastatic CRC treated with a combination of bevacizumab, irinotecan, fluorouracil, and leucovorin.

In the present study, either modified IFL regimen (modified irinotecan, fluorouracil and leucovorin, mIFL) alone or in combination with bevacizumab was used to treat patients with metastatic colorectal cancer (CRC). Treatment efficacy was assessed using coupled tomography imaging diagnosis. The toxicity accompany with treatment was evaluated, as well as T cell receptor (TCR) repertoire before and several cycles after therapy was dynamically monitored by analyzing the complementarity-determining region 3 (CDR3) length distribution within CD4(+) and CD8(+) T cell subsets. The degrees of normalization of the T cell repertoire in CRC patients treated with the two methods were compared. The results showed that mIFL combined with bevacizumab was more effective in treating patients with metastatic CRC, and was accompanied by an increase in side effects such as proteinuria and hematuria. An even more restricted CDR3 profile in patients with metastatic CRC compared with healthy control has been detected. A prominent usage of TCR beta chain variable (BV) gene BV12 and BV16 families within the CD4(+) T cell subset and BV19 and BV21 families within the CD8(+) T cell subset have been found before treatment. Moreover, CD8(+) T cells showed more restricted patterns than CD4(+) T cells, especially in patients before treatment. For patients with stable disease (SD) or partial remission (PR) after treatment, a less restricted CDR3 profile in post-treatment compared with pre-treatment has been found, but the opposite result was observed for patients with progressive disease (PD). The less restricted CDR3 pattern suggested a trend toward normalization of the TCR repertoire. The normalization of TCR repertoire significantly increased in patients treated with mIFL in combination with bevacizumab, but slightly in patients treated with mIFL alone. The results demonstrate a positive correlation between post-therapy TCR repertoire normalization and remission of metastatic CRC.

[Evaluation of tumor angiogenesis using microbubbles conjugated with RGD peptides and contrast enhanced ultrasound].

OBJECTIVE: To assess the feasibility of usage of microbubbles conjugated with RGD peptides and contrast enhanced ultrasound (CEU) in detection of tumor angiogenesis. METHODS: Lipid microbubbles (MB) were prepared, and the RGD peptides were covalently conjugated to the lipid shell of MB (MB(RGD)). Six nude mice with tumor created by dorsal inoculation of HepG2 tumor cells were used as the test group. Six nude mice without tumor were served as the control group. 10 minutes after bolus injection of MB and MB(RGD) randomly (30 min interval) via a tail vein catheter, CEU was performed on the tumors of the test group and the thigh skeletal muscles of control group. The video intensity (VI) of tumors and the skeletal muscles were measured. The tumors and the skeletal muscles were harvested for immunohistochemical examination. RESULTS: Only a slight contrast enhancement of the tumor was seen with MB, and the VI was 5.33 ± 1.71. While a remarkable enhancement of the tumor was observed after injection of MB(RGD). The VI was up to 17.03 ± 3.58, 3.18 folds higher as compared with that obtained by injection of MB (P < 0.05). As expected, there were no obvious contrast enhancement of the skeletal muscles with both MB(RGD) and MB. There was a high expression of αvß3-integrin in tumor neovascular endothelium, however, no apparent expression of αvß3-integrin was observed in the skeletal muscle vascular endothelium. CONCLUSION: CEU with MB(RGD) can be used to effectively evaluate the angiogenesis of tumors, and it may greatly contribute to the early judgement of the nature of tumor.

Trastuzumab plus docetaxel and capecitabine as a first-line treatment for HER2-positive advanced gastric or gastroesophageal junction cancer: a phase II, multicenter, open-label, single-arm study.

Gastric cancer (GC) is the second most common cancer in China. The ToGA study showed that trastuzumab in combination with fluoropyrimidine plus cisplatin prolonged overall survival (OS) in patients with human epidermal growth factor receptor 2 (HER2)-positive advanced GC (AGC). However, some patients may not be able to receive this regimen. We conducted a clinical study to evaluate the efficacy and safety of trastuzumab in combination with docetaxel+capecitabine (DX) in patients with HER2-positive AGC. This phase II, multi-center, open-label, single arm study enrolled patients with HER2-positive AGC who had not received prior treatment for metastatic disease. Patients were treated with a regimen of trastuzumab (8 mg/kg loading dose followed by 6 mg/kg, day 1), capecitabine (1000 mg/m2 twice daily, days 1-14) and docetaxel (60 mg/m2, day 1 for 6 cycles) every 3 weeks. The primary endpoint was progression-free survival (PFS) and the secondary endpoints were objective response rate (ORR), OS and safety profiles. Sixty-seven patients with AGC were enrolled from 14 centers. 64 were included in the full analysis set (FAS). The median PFS was 8.1 months (95% confidence interval [CI]: 5.6-12.8) and the median OS was 20.9 months (95% CI: 15.1-33.0). Response was evaluated in 59 patients. The ORR was 67.8%. The most common adverse events of Grade ≥3 were neutropenia, leukopenia, hand-foot syndrome, febrile neutropenia and anemia. We concluded that combination treatment with trastuzumab and DX was well-tolerated and highly effective in patients with HER2-positive AGC, and may offer an alternative to current treatments.

Clinical study of Photofrin photodynamic therapy for the treatment of relapse nasopharyngeal carcinoma.

OBJECTIVES: To evaluate the clinical response and adverse effects of Photofrin photodynamic therapy (PDT) in patients with relapse nasal pharyngeal cancer. METHODS: Thirty patients with relapse nasal pharyngeal cancer were randomly divided into PDT group and chemotherapy group. In PDT group, patients received intravenous administration of Photofrin (2mg/kg b.w.) and 48h later light (200-300J/cm) was delivered by a 630nm diode laser through a cylinder diffuser under endoscopic assistance. Two days after PDT the necrotic tissues were removed and newly identified sites were subject to another round of light irradiation. In chemotherapy group, the routine cisplatin and 5-fluorouracil (DDP/5-FU) remedy was used. Endoscopic and radiological examinations were performed during 6 months follow-up. The Karnofsy Score was used to evaluate the quality of life. RESULTS: The local response and nasal cavity obstruction remission rate in PDT group were better than that in chemotherapy group. The Karnofsy Score was also improved in PDT group. CONCLUSION: This pilot study suggests that Photofrin PDT is effective and safe in treatment of advanced nasal pharyngeal cancer and management of nasal obstruction.

Metastasis-associated in colon cancer-1 in gastric cancer: Beyond metastasis.

Metastasis-associated in colon cancer-1 (MACC1) is an oncogene that was first identified in colon cancer. The upstream and downstream of MACC1 form a delicate regulatory network that supports its tumorigenic role in cancers. Multiple functions of MACC1 have been discovered in many cancers. In gastric cancer (GC), MACC1 has been shown to be involved in oncogenesis and tumor progression. MACC1 overexpression adversely affects the clinical outcomes of GC patients. Regarding the mechanism of action of MACC1 in GC, studies have shown that it promotes the epithelial-to-mesenchymal transition and accelerates cancer metastasis. MACC1 is involved in many hallmarks of GC in addition to metastasis. MACC1 promotes vasculogenic mimicry (VM) via TWIST1/2, and VM increases the tumor blood supply, which is necessary for tumor progression. MACC1 also facilitates GC lymphangiogenesis by upregulating extracellular secretion of VEGF-C/D, indicating that MACC1 may be an important player in GC lymphatic dissemination. Additionally, MACC1 supports GC growth under metabolic stress by enhancing the Warburg effect. In conclusion, MACC1 participates in multiple biological processes inside and outside of GC cells, making it an important mediator of the tumor microenvironment.

[Killing effect of Photofrin-Herceptin immunoconjugate on tumor cell lines in vitro].

OBJECTIVE: To study the specific killing effect of the Photofrin-Herceptin immunoconjugate on the tumor cells expressing HER-2. METHODS: Photofrin (porfimer sodium) was covalently coupled to Herceptin (trastuzumab) via [1-ethyl-3-(3-dimethyl-aminopropyl)carbodiimide hydrochloride](EDCI) , and the killing effect of this conjugate was evaluated by means of MTT assay on SK-BR-3, MCF-7 and A549 cell lines expressing HER-2 at different levels. RESULTS: The conjugate had weaker immunoactivity than Herceptin, but possessed stronger killing effect on the cell lines expressing HER-2 than Photofrin, Herceptin, as well as the mixture of Photofrin and Herceptin (P<0.05). In the cell lines negative for HER-2 expression, the killing effect of the conjugate was similar to that of Photofrin, Herceptin, and their mixture. CONCLUSION: Photofrin-Herceptin immunoconjugate can specifically kill HER-positive cells in vitro.

[Effect of Herceptin combined with Taxol on patients with Her-2/neu overexpressing metastatic breast cancer].

OBJECTIVE: To investigate the efficacy and toxicity of recombinant humanized anti-Her-2/neu antibody (Herceptin) and Taxol for patients with Her-2/neu overexpressing metastatic breast cancer. METHODS: Sixty patients with Her-2/neu overexpressing metastatic breast cancer were investigated. Of the 60 cases, 22 were treated with Herceptin and Taxol and 38 with Taxol and doxorubicin. RESULTS: The total response rate (RR) of Herceptin and Taxol was 68.2%, and that of Taxol and doxorubicin was 44.7%. The RR of patients with Her-2/neu(+++) was 75%, while that of patients with Her-2/neu(++) was 50%. The major adverse effects were gastro-intestinal tract reactions, myopathy, bone marrow suppression and alopecia. CONCLUSION: The treatment with Herceptin and Taxol is effective and safe for patients with Her-2/neu overexpressing metastatic breast cancer. The therapeutic effect is related to the degree of Her-2/neu overexpression.

[Investigation of short-term therapy results for radiofrequency ablation by positron emission tomography].

OBJECTIVE: To explore the possibility of evaluating the short-term therapeulic effect of radiofrequency ablation (RFA) in cancer treatment by positron emission tomography. METHODS: The radioactivity intensity of the tumor was detected by PET before and after RFA. RESULTS: Radioactivity blank was observed in all 33 cases with 54 lesions, indicating the elimination of the tumor was destroyed. CONCLUSION: PET is of great significance in evaluating the short-term effect of RFA.

[Clinical study of Photofrin photodynamic therapy for advanced cancers].

OBJECTIVE: To evaluate the clinical efficacy and adverse effects of Photofrin photodynamic therapy (PDT) in patients with advanced cancers. METHODS: Forty patients with advanced cancers in stage IV with lumen obstruction, who failed to respond positively to other treatment regimens, received intravenous administration of Photofrin as the photosensitizer at the dose of 2 mg/kg.b.w. 48 h before PDT by 630 nm light (DIOMED) delivered through cylinder diffusing tip quartz fibers that passed through the biopsy channel of a flexible endoscope. PDT endoscopy was repeated, the necrotic tissue removed and, if necessary, the primary sites and other newly identified sites were subjected to a second exposure 8 h later. Two days after the second exposure, endoscopy was again performed and the necrotic tissue removed. Endoscopy was repeated one month after PDT and periodically thereafter as needed to treat symptomatic residual tumor. RESULTS: The total rate of response to the treatment was 74% in these patients, and the rate of lumen obstruction due to the tumors decreased from 90% to 10% after PDT, with significantly improved Karnofsky performance score. CONCLUSION: Photofrin PDT is effective and safe in the treatment of advanced cancer, which may relieve lumen obstruction and improve patient quality of life.

Association of progesterone receptor and HER2 expressions with the survival time of patient with breast cancer.

OBJECTIVE: To investigate the relationship between the survival time and the expressions of progesterone receptor (PR) and HER2 in the tumor tissues of patients with breast cancer, thereby to explore the possibility of using the 2 indices as the indicator for the prognosis of the patients. METHOD: Immunohistochemical staining for PR and HER2 was performed on paraffin embedded tumor specimens obtained from 185 patients with breast cancer, and follow-up studies were conducted and the survival time of the patients recorded. RESULTS: In the 120 patients (64.9%) with complete followed-up data, 28 patients died (15%), with the record of the other 65 patients (35.1%) not available for this study. Immunohistochemical detection found that the positivity rates for HER2 and PR were 57.8% and 62.7% respectively in the total specimens examined, and HER2 and PR showed significant difference in their respective associations with the survival rate of the patients (P<0.01). Specifically, HER2 was inversely associated with the survival rate, to which PR was positively related. The univariant analysis suggested the hazards posed by HER2 expression to reduce the overall survival of the patients (P<0.01, OR=1.566), while PR, playing the protective role, was associated with a significantly longer overall survival time (P<0.01, OR=0.547). In the multivariate analysis, however, only PR expression was of significant prognostic value in the patients (P 0.012). CONCLUSION: Both HER2 and PR can be used as independent indicators for the prognosis of patients with breast cancer, and high HER2 expression may indicate poor prognosis, while PR is associated with a longer survival time and a higher survival rate.

[Establishment of BGC-823/pBaBb-puro-MACC1 gastric cancer cell line stably expressing MACC1 and its tumor-related gene expression profiles].

OBJECTIVE: To establish a gastric cancer cell line with stable expression of metastasis-associated in colon cancer 1 (MACC1) and detect the changes in tumor-related gene expression profiles for investigating the possible regulation mechanisms between MACC1 and the differentially expressed genes. METHODS: The full-length MACC1 cDNA was amplified from human embryonic kidney 293FT cells and cloned into the pBaBb-puro vector. The recombinant pBaBb-puro-MACC1 expression vector, after identification with restriction enzyme digestion, was transfected into 293FT cells, and the expression of fluorescent reporter gene was observed. pBaBb-puro-MACC1 vector was transfected into human gastric cancer BGC-823 cell line to establish BGC-823/pBaBb-puro-MACC1 cell line stably expressing MACC1. Quantitative RT-PCR and Western blotting were used to detect MACC1 expression in both BGC-823/pBaBb-puro-MACC1 and control BGC-823 cells. High-throughout cDNA microarray was used to screen the effects of MACC1 on the gene expression profiles of gastric cancer cells. RESULTS: The recombinant pBaBb-puro-MACC1 plasmid was successfully constructed and verified by PCR and sequencing. BGC-823/pBaBb-puro-MACC1 cells showed significantly increased MACC1 mRNA expression as compared with the control cells. The results of cDNA microarray identified 33 up-regulated and 24 down-regulated genes in the cells after MACC1 transfection involved were in various cellular functions. CONCLUSION: The established BGC-823/pBaBb-puro-MACC1 gastric cancer cell line show some important molecular changes caused by MACC1.

KRAS and PIK3CA but not BRAF genes are frequently mutated in Chinese cholangiocarcinoma patients.

Cholangiocarcinoma (CCA) is a rare but lethal malignancy arising from the biliary tract epithelium. It has a poor prognosis largely due to the difficulties of early diagnosis and the lack of effective therapies. It is thus imperative to develop new and effective treatments for CCA, which depends heavily on the mechanistic understanding of the disease. Previous studies have suggested that somatic mutations in KRAS, BRAF, and PIK3CA genes are frequently found in several types of human cancers including colon, breast, and lung carcinomas as well as CCA. Yet, the frequency and the involvement of these oncogenic mutations in CCA in Chinese population have not been investigated. In this study, we evaluated the hotspot mutations of KRAS, BRAF, and PIK3CA genes in 34 Chinese CCA patients. Sequencing analysis revealed 13 (38.2%) and 11 (32.4%) patients bearing KRAS and PIK3CA mutations, in which two (5.9%) of them harbored both KRAS and PIK3CA mutations. Surprisingly, no BRAF mutation was detected in all 34 CCA samples. Our findings indicate that somatic mutations in KRAS and PIK3CA but not BRAF oncogenes are closely associated with the development of CCA in Chinese population and provide new potential targets for future therapeutic treatments of the disease.

Local recurrence of hepatocellular carcinoma after radiofrequency ablation.

A 51-year-old Chinese male with a 20-year history of hepatitis B was diagnosed with hepatocellular carcinoma in the right anterior portion of the liver, sized 3.5 cm × 3.2 cm, and was treated with radiofrequency ablation (RFA) on December 18, 2001. The patient did not receive antiviral therapy for hepatitis B virus after RFA. The treated lesion reduced gradually and reached its minimum size of 1.7 cm × 1.5 cm seven years later on November 18, 2008. However computed tomography findings revealed that a recurrence lesion of 6.0 cm × 4.8 cm which was histologically confirmed overlapped the previous treated lesion at the 8th year on December 3, 2009. Although recurrence at 8 years after curative RFA is a rare event, such a possibility must be kept in mind. To find and treat the recurrence lesion promptly, long-term and close monitoring is warranted after RFA. Meanwhile, the recurrence-prevention therapy is as important as close monitoring for those patients with a history of hepatitis B.

[Short-term therapeutic effect and safety of endostar combined with XELIRI regimen in the treatment of advanced colorectal cancer].

OBJECTIVE: To evaluate the short-term efficacy and toxicity of endostar in combination with XELIRI as the second-line treatment for advanced colorectal cancer. METHODS: Twenty-one patients with advanced colorectal cancer were treated with intravenous infusion of endostar (15 mg/day for 14 consecutive days) and irinotecan (250 mg/m(2), single dose on the first day), and oral administration of capecitabine (1.0 mg/m(2), twice daily for 14 days), and the treatment cycle was repeated every 21 days. The efficacy and toxicity of the treatments were evaluated according to RECIST and NCI-CTCAE3.0 standard, respectively. RESULTS: The overall response rate was 9.5% in these patients, with a median time to progression (mTTP) of 3.9 months. The main adverse effects associated with the treatment included leucopenia, nausea/vomiting and peripheral neuritis. CONCLUSION: Endostar combined with XELIRI is effective and safe as the second-line treatment for advanced colorectal cancer, and further clinical investigation is warranted.

[Ultrasound-mediated microbubble destruction enhances LMP-1 gene transfection into dendritic cells in vivo].

OBJECTIVE: To investigate the transfection efficiency and the optimal conditions of delivering latent membrane protein-1 (LMP-1) gene to dendritic cells (DCs) by ultrasound exposure combined with contrast agent. METHODS: Human DCs were cultured in vivo and transfected with the recombinant plasmid pEGFP-C3-LMP1 under varying conditions including ultrasound intensities, exposure time and microbubble contrast agent concentration. The transfection efficiency was assessed by fluorescent microscopy and flow cytometry, and the cell viability by trypan blue exclusion test. RESULTS: An exposure time of 60 s at MI 1.0 with a microbubble contrast agent concentration of 20% resulted in the optimal effect of delivering the recombinant plasmid pEGFP-C3-LMP1 into the DCs, with a transfection efficiency of (14.37∓2.12)%. Over 90% of the transfected cells were viable after the transfection. CONCLUSION: Microbubble contrast agent combined with ultrasound exposure can enhance the delivery of recombinant plasmid pEGFP-C3-LMP1 into the DCs.