In particular, breast cancer and hepatocellular carcinoma (HCC) exhibit accelerated pathological progression in patients with metabolic syndrome. In this thesis, we investigated the coupling factors involved in the association of metabolic dysfunctions with CLD and breast cancer and how they are regulated and how they contributed to the pathophysiology of the diseases. It has previously been documented to be excessively generated in adipose tissue under obese conditions, which in turn contributes to the pathological process of the metabolic diseases.
The effects of the compounds in combination therapy with cisplatin were partially mediated by suppression of the epithelial-mesenchymal transition (EMT) of breast cancer cells. In addition, we suggest that the metabolic status of patients should be taken into account in order to achieve an optimal therapeutic strategy for patients with comorbidities. Crosstalk of Notch and HER3 signaling via Nrg1 promotes breast cancer progression in diabetic patients.
Introduction
Considering current trends in obese population, the incidence and mortality of HCC is expected to be significantly increased. Those epidemiological associations between cancer and type 2 diabetes are attributed to various factors (ie, hyperinsulinemia, hyperglycemia, or dyslipidemia). The NICD-RBPJ complex contributes to turn on the transcriptional program to activate Notch target genes, such as the HES or HEY.
Among the variants, type I isoforms include Neu differentiation factor (NDF), hereregulin, (HRG) and acetylcholine receptor-inducing activity (ARIA), all of which have an immunoglobulin-like domain. Nominative identification (ie, NRG1-type1 β1a) is determined by variations in structure; C-terminus of the EGF domain (α, β and y) and in linker (1 to 4) and in the intracellular domain (a to c). Verminoside (Vms) is extracted from leaves of NC13, one of the main bioactive constituents of NC13.
Materials and Experimental Method
All breast cancer cells were maintained in low-glucose DMEM for at least 3 days and then transferred to high-glucose (4.5 g/L) DMEM until the indicated time points. Cancer cells were treated with lentiviral and polybrene media, and infected cells were selected by puromycin treatment. After the indicated treatments, cells were washed twice with PBS and fixed in a PFA (4%) for 30 min at 25℃.
Apoptotic cells were detected using TUNEL assay kit (Roche St. Louis, MO, USA), according to the manufacturer's protocols. After HG treatment, the cells were washed twice with PBS and then fixed in a PFA (4%) in PBS for 30 min. Breast cancer cells were plated overnight in a 96-well culture dish (2000 cells/well) and then treated with siRNA or DAPT.
Results
Part 1. COL6A3-derived endotrophin links reciprocal interactions among hepatic cells in the pathology of chronic liver disease
After administration of doxycycline (Dox) to Alb-ETP. mouse model, endotrophin expression could be induced in liver tissue. Endotrophin levels were increased in the liver tissue of CCl4-treated mice, which were further increased in Alb-ETP mice (Figure III-1-4C). Hepatic apoptosis was increased in the liver tissues of the Alb-ETP group of mice compared with the control, as assessed by cleaved caspase 3 staining (Figure III-1-4E).
Similarly, fibrosis and inflammation were augmented in liver tissue samples from Alb-ETP group, as assessed by αSMA, Masson's trichrome C and Mac2 staining (Figure III-1-4F-H). Most of the mice showed tumor development in liver tissue, where the Alb-ETP mice had increased tumor size and number compared to control (Figure III-1-5A). Cancer incidence was slightly increased in Alb-ETP mice and they had a significant increase in the number of primary tumor nodules in liver tissues (Figure III-1-5C-D).
Immunostaining of liver tissue showed that Alb-ETP mice had an increased proportion of Ki-67 positive cells, especially near tumor lesions, compared to control mice (Figure III-1-5E). Consistently, Alb-ETP mice had increased serum levels of AFP, a marker of liver cancer (Figure III-1-5F). There were increased expressions of several markers of inflammation and fibrosis in the liver tissues of Alb-ETP mice compared with those of controls (Figure III-1-5J,K).
Immunostaining of αSMA in liver tissue also showed that Alb-ETP mice had elevated αSMA-positive surface area compared to control mice (Figure III-1-5L). Similarly, Alb-ETP mice had aggravated liver fibrosis, apoptosis, and increased periportal area compared with control mice (Figure III-1-7A-E). Alb-ETP mice had increased JNK1 phosphorylation and cleaved caspase 3 levels compared to control mice (Figure III-1-5M).
Alb-ETP and control mice were treated with a single injection of DEN (25 mg/kg) on postnatal day 14 and fed a dox diet (200 mg/kg).
Part 2. Notch and HER3 signaling crosstalk via Nrg1 promotes breast cancer progression in patients with diabetes
Consistent with the status of active histone marks, chromatin accessibility to the Nrg1 enhancer (+200 bp) was also increased by HG treatment in breast cancer cells, indicating that HG induced an open chromatin landscape within the Nrg1 enhancer region. -J) ChIP-qPCR analysis of time-course binding occupancy of P300, CBP, SETD1A and HDAC1 at the Nrg1 enhancer in HG-treated Met1 cells. Thousands of candidates were enriched in the Nrg1 enhancer sequence after HG treatment, and five candidates were selected after a literature search (Figure III-2-5B).
NICD was recruited to the Nrg1 enhancer region in HG-treated Met1 cancer cells (Figure III-2-6C). Since putative RBPJ-binding sequences (TGGGAA) were found in the Nrg1 enhancer (Figure III-2-6D), we analyzed whether RBPJ is recruited to the Nrg1 enhancer by HG. After HG treatment, the activity in the Nrg1 enhancer and the 1 kb region of the Nrg1 promoter both increased.
Luciferase activity was elevated by RBPJ and further increased in the presence of the Nrg1 enhancer, suggesting that RBPJ may induce activation of the Nrg1 enhancer. HG-treated Met1 cells were treated with DAPT and the occupancy of the NICD-RBPJ junction with various chromatin remodelers, including P300, SETD1A, and HDAC1, at the Nrg1 enhancer was analyzed. HG-induced recruitment of NICD-RBPJ, P300, CBP, and SETD1A to the Nrg1 enhancer was reduced, whereas HG-induced HDAC1 dissociation was suppressed by DAPT (Figure III-2-9C-H).
Similarly, the two active histone modifications were enriched upon Notch activation (Figure III-2-10L-N), suggesting that NICD overexpression is sufficient to establish active histone marks in the Nrg1 enhancer and drive Nrg1 overexpression . -N) Binding enrichment of (L) Rbpj, (M) H3K4me1 and (N) H3K27ac within the Nrg1 enhancer was increased by NICD overexpression in 4T1 cells, as determined by ChIP-qPCR. This is consistent with previous reports that AP1 can drive transcriptional activity of the Nrg1 enhancer5.
The chromatin of the NRG1 enhancer region was also more accessible in the high-NOTCH group than in the low-NOTCH group, based on ATAC sequencing results (Figure III-2-14C). Since the NRG1-HER3 axis is involved in resistance to HER2-targeted therapy, the impact of NRG1 levels was investigated in HER2-positive breast cancer patients instead of the entire TCGA breast cancer cohort. The NRG1 enhancer regions are highlighted as light blue, showing that the enhancer is well conserved.
Part 3. Natural product derived Verminoside confers cisplatin sensitivity in metastatic breast cancer
Tables
- Discussion
GTT TGT GGT AGA GAA CAG AAA ACA G Forward GCA TGT AAG ATC TTG GTA CTT TCC C Reverse human Nrg1 amplifier.
Part 1. COL6A3-derived endotrophin links reciprocal interactions among hepatic cells in the pathological process of liver cancer
Crosstalk of Notch and HER3 signaling via Nrg1 promotes breast cancer progression in diabetic patients, diabetic patients. The therapeutic potential of the metabolic status of breast cancer patients has been largely ignored, as therapeutic results have been unsatisfactory. Approximately 35% of patients with HER2-positive breast cancer initially responded to trastuzumab, of which only 30% did not show progression within one year [60].
The deleterious impact of diabetes on the clinical outcomes of HER2-positive breast cancer treatment with trastuzumab was reported [61, 62]. Considering the applications of the Notch inhibitors in cancer and diabetes, Notch-targeted drugs are expected to be beneficial, especially in diabetic patients with cancer. Because Notch activity is increased in HG-treated cancer cells and tissues from HER2-positive breast cancer patients with hyperglycemia, we postulated that hyperglycemia-driven NOTCH activation in breast cancer cells exacerbated resistance to HER2-targeted therapy.
Furthermore, NRG1 overexpression predicted poor outcome in hyperglycemic HER2-positive cancer patients, but not in HER2-negative patients. One example of successful multidrug therapy is the administration of cisplatin in combination with gemcitabine or docetaxel, which are currently the first-line chemotherapy protocols for patients with metastatic breast cancer [71, 72]. National Cancer Institute guidelines describe adjuvant therapy as an additional intervention for cancer patients who have received primary therapeutic treatment to reduce the risk of cancer recurrence.
The target compounds were screened based on cytotoxic effects on the breast cancer cells, with the synergistic (or additive) cytotoxic effect exerted only when used in combination with cisplatin. We propose that diabetes status and the NOTCH-NRG1-HER3 axis can be used as prognostic markers to predict responses to HER2-targeted therapy, offering a unique therapeutic strategy for the treatment of breast cancer patients with diabetes. Numerous studies reported the promising effects of anticancer drugs on metastasis in breast cancer models [83–85], and furthermore, several drugs for metastatic breast cancer patients are in phase III clinical trials, including sacituzumab and govitecan-hziy [86].
Various applications of combination therapy have been investigated to overcome the limits of conventional chemotherapy, and our results suggested that Vms could be developed as a chemoadjuvant for metastatic breast cancer patients.
ⅥI. References
Chang et al., "Persistent JNK1 activation is associated with altered histone H3 methylations in human liver cancer," Journal of hepatology, vol. Vasconcelos-Dos-Santos et al., "Hyperglycemia exacerbates malignancy in colon cancer through the hexosamine biosynthetic pathway," Oncogenesis, vol Minning et al., “Exploring breast carcinogenesis through integrative genomics and epigenomic analyses,” Int J Oncol, vol.
Stadler et al., “DNA-binding factors shape the mouse methylome in distal regulatory regions,” Nature, vol. Jin et al., “Aberrant expression of SETD1A promotes the survival and migration of estrogen receptor α-positive breast cancer cells,” International journal of cancer, vol. Yang et al., “NRG1-dependent activation of HER3 induces primary resistance to trastuzumab in HER2-overexpressing breast cancer cells,” International Journal of Oncology, vol.
Lee et al., “Akt-dependent metabolic reprogramming regulates tumor cell histone acetylation,” Cell Metabolism , vol. Lee et al., “Diabetes as a prognostic factor in HER-2 positive breast cancer patients treated with targeted therapy,” Breast Cancer , vol. Abravanel et al., "Notch promotes recurrence of dormant tumor cells after HER2/neu-targeted therapy," The Journal of clinical study, vol.
Zhou et al., "Notch signaling pathway: Architecture, disease, and therapeutics," Signaltransduktion og målrettet terapi, vol. Zhang et al., "Cisplatin and gemcitabin as the first line therapy in metastatic triple negative breast cancer," International journal of cancer, vol. Vassilomanolakis et al., "First-line kemotherapy with docetaxel and cisplatin in metastatic breast cancer," The Breast, vol.
Bardia et al., “Sacituzumab govitecan-hziy in refractory metastatic triple-negative breast cancer,” New England Journal of Medicine , vol.
ⅤIII. Acknowledgement
