Rat Breast Tumor Research Articles

The effect of non‐contact electro capacitive cancer therapy on DMBA‐induced rat breast tumor angiogenesis

Alternating Current‐Electric Field (AC‐EF) generated by non‐contact Electro Capacitive Cancer Therapy (ECCT) can inhibit breast tumor growth. However, its effect on breast tumor angiogenesis remains unclear. Since angiogenesis is involved in normal physiology and tumors, it is crucial to investigate the effect of ECCT on normal and breast tumor angiogenesis. Samples consisting of rat breast normal tissue and breast tumors were obtained from the biobank, with tumors induced by 7,12‐dimethylbenz (α) anthracene (DMBA) at 20 mg/kg BW 10 times over five weeks. Meanwhile, ECCT exposure of 150 kHz and 18 Vpp was conducted for 21 days at 10 hours/day. The qPCR method was used for gene expression analysis, while immunohistochemistry used antibody anti‐Vegfr2 that was used to detect Vegfr2 protein expression. Data were analyzed using one‐way ANOVA and t‐tests performed with GraphPad Prism ver.9.5.1 software. The results revealed no impact of ECCT exposure on normal breast tissue angiogenesis. Interestingly, there was a significant increase in the number of blood vessels following the upregulation of Vascular Endothelial Growth Factor Receptor‐2 (Vegfr2) as opposed to its primary signal, Vascular Endothelial Growth Factor‐A (Vegfa). Furthermore, gene expression of Hypoxia Inducible Factor‐1α (Hif1α) and Specificity Protein‐1 (Sp1) was similar to that of the control group, suggesting that Vegfr2‐dependent angiogenesis regulates ECCT‐treated breast tumor angiogenesis.

Ultrasound mediated sonochemical internalization (SCI): overcoming the depth limitations of light-based therapies

SignificanceFocused ultrasound activation of sonosensitizers i.e. sonodynamic therapy (SDT) is an alternative to light-activated photodynamic therapy (PDT). Sonochemical internalization (SCI), is comparable to light based photochemical internalization (PCI) but without the intrinsic drawback of limited penetration depth of light in biological tissues including the skull. ApproachSCI experiments were performed both in vitro on monolayers and in vivo, in a rat breast tumor model, using the sononsensitizer AlPcS2a and the drug bleomycin (BLM). Resultsin vitro; Growth inhibition of tumor cells in monolayer was significantly greater than that seen with either BLM or SDT acting alone. Similar results were also seen in vivo, with significant reduction in breast tumor development following BLM-SCI ConclusionThe synergist effect of SCI, employing relatively low drug concentrations and focused ultrasound power levels, would reduce the problems of nonspecific tissue damage caused by thermal effects at high power and unwanted systemic drug side effects.

Exploring the Protective Effect against 7,12-Dimethylbenz[a]anthracene-Induced Breast Tumors of Palmitoylethanolamide.

Breast cancer remains a global health burden, and the need for effective therapies is of chief importance. The current study explored the in vivo chemoprotective activity of palmitoylethanolamide (PEA) against 7,12-dimethylbenz[a]anthracene (DMBA)-induced breast tumor in rats. Results of noninvasive photoacoustic imaging showed real-time progression in the tumor area and volume in DMBA-induced rats, while there was a reduction in tumor area and volume in PEA-treated tumor-bearing rats. The increase in the average oxygen saturation (sO2 %) and decrease in the average total hemoglobin (HbT %) indicated the PEA-mediated attenuation of hypoxia-induced neovascularization in DMBA-induced rats. Histopathological investigations confirmed the efficacy of PEA in mitigating breast carcinoma, hepatotoxicity and nephrotoxicity driven by DMBA. Moreover, PEA-mediated alterations in the metabolic activity of the tumor microenvironment were evidenced by decreased glucose and lactate dehydrogenase enzyme level in the blood plasma and mammary tissue. PEA also maintained the redox balance by inhibiting nitric oxide level, reducing malondialdehyde (a product of lipid peroxidation), and increasing the level of antioxidant enzyme reduced glutathione. PEA altered the expression of apoptosis-related genes (BAX, P53,BCL-XL, CASPASE-8, and CASPASE-9) and induced the activity of Caspase-3 protein in the mammary tissue of tumor-bearing rats, indicating its apoptosis inducing ability. Taken together, the findings of this study suggest that PEA may have a protective effect against DMBA-induced breast tumors.

Hexyl-pentynoic acid serves as a novel radiosensitizer for breast cancer by inhibiting UCHL3-dependent Rad51 deubiquitination

ObjectiveTo investigate the effects and underlying mechanism of 2-hexyl-4-pentynoic acid (HPTA), a derivative of valproic acid (VPA), on radiotherapy in breast cancer. MethodsMCF7 cells and 7,12-dimethylbenz-[α]-anthracene (DMBA)-induced transformed human normal breast cells (MCF10A–DMBA cells) were irradiated with 8 ​Gy X-rays. For both cells there were four groups: control, valproic acid (VPA)/HPTA, IR, and VPA/HPTA+IR groups. MTT and clonogenic survival assays were performed to assess cell proliferation, and comet assay was performed to evaluate DNA damage. Protein expression of γH2AX, 53BP1, Rad51, and BRCA1 was examined via immunofluorescence and immunoblotting. Cycloheximide chase and ubiquitination experiments were conducted to determine Rad51 ubiquitination. In vivo experiments involved a rat model of DMBA-induced breast cancer, with four fractionated doses of 2 ​Gy. Tumor tissue pathological changes and γH2AX, Rad51, and UCHL3 expression levels were measured by hematoxylin-eosin staining, immunohistochemistry, and immunoblotting. ResultsCompared with the IR group, 15 ​μmol/L HPTA reduced the cell proliferation ability of irradiated MCF7 cells (t=2.16, P<0.05). The VPA/HPTA+IR group exhibited significantly increased DNA double-strand breaks relative to those in the IR group (VPA+IR vs. IR, t=13.37, P<0.05; HPTA+IR vs. IR, t=8.48, P<0.05). Immunofluorescence and immunoblotting experiments demonstrated that the VPA/HPTA+IR group displayed significantly increased cell foci formation, γH2AX and 53BP1 protein expression levels compared to the IR group [(γH2AX: VPA+IR vs. IR, t=8.88, P< 0.05; HPTA+IR vs. IR, t=8.90, P< 0.05), (53BP1, VPA+IR vs. IR, t=5.73, P< 0.05; HPTA+IR vs. IR, t=6.40, P< 0.05)]. Further, Rad51 expression was downregulated (VPA+IR vs. IR, t=3.12, P< 0.05; HPTA+IR vs. IR, t ​= ​2.70, P<0.05), and Rad51 inhibition effectively counteracted HPTA-induced radiosensitization. Ubiquitination detection further verified that HPTA inhibits Rad51 expression via UCHL3-dependent Rad51 deubiquitination. In vivo study results showed that 20 ​mg/kg HPTA significantly enhanced the radiosensitivity of breast tumors in rats by inhibiting Rad51 expression. ConclusionsHPTA is a highly effective radiosensitizer that enhances the radiotherapeutic efficacy of breast cancer treatment through UCHL3-dependent deubiquitination of Rad51.

Obesity Associated with Prediabetes Increases the Risk of Breast Cancer Development and Progression-A Study on an Obese Rat Model with Impaired Glucose Tolerance.

Patients with comorbidities of obesity and diabetes are recognized to be at high risk of breast cancer development and face worse breast cancer outcomes. Though several reports showed the reinforced link between obesity, diabetes, and prediabetes with breast cancer, the underlying molecular mechanisms are still unknown. The present study aimed to investigate the underlying molecular link between increased risks of breast cancer due to coincident diabetes or obesity using a spontaneous obese rat model with impaired glucose tolerance (WNIN/GR-Ob rat). A single dose of solubilized DMBA suspension (40 mg/kg body weight) was orally administered to the animals at the age of 60 days to induce breast tumors. The tumor incidence, latency period, tumor frequency, and tumor volume were measured. Histology, immunohistochemistry, and immunoblotting were performed to evaluate the tumor morphology and expression levels of signal molecules. The development of mammary tumors in GR-Ob rats was characterized by early onset and shorter latency periods compared to control lean rats. While 62% of obese rats developed breast tumors, tumor development in lean rats was only 21%. Overexpression of ER, PR, Ki67, and p53 markers was observed in tumor tissues of obese rats in comparison with lean rats. The levels of the hallmarks of cell proliferation and angiogenesis involved in IGF-1/PI3K/Akt/GSK3β/β-catenin signaling pathway molecules were upregulated in obese rat breast tumors compared to lean rats. Furthermore, obesity with prediabetes is associated with changes in IGF-1 signaling and acts on PI3K/Akt/GSK3β/β-catenin signaling, which results in rapid cell proliferation and development of breast tumors in obese rats than the lean rats. These results indicate that tumor onset and development were faster in spontaneous obese rat models with impaired glucose tolerance than in their lean counterparts.

Inhibition of Dimethylbenz[a]anthracene-Induced Breast Tumors in Rats by Soy Protein is Mediated by Downregulation of MAPK/AP-1 Signaling

We reported earlier that replacement of casein by soy protein in the diets of rats exposed to the carcinogen, dimethylbenz[a]anthracene (DMBA), not only delayed the initiation of breast tumor but also had a protective effect against the development of aggressive tumor. The aim of this study was to elucidate the molecular mechanism by which soy protein offers these beneficial effects. Tumor was developed by gavage administration of single dose of 80 mg/kg of DMBA into 50-day old female rats, maintained on a standard AIN-76A diet containing either casein or soy protein. After ~120 days of DMBA administration, we evaluated the role of MAPK phosphorylation and subsequent AP-1 activation on the differential effects of soy protein and casein on the development of aggressiveness and progression of DMBA-induced breast tumor and determined if soy protein controls MMP-9 and uPAR expression by modifying AP-1 activity. The present study demonstrates that the beneficial effect of soy protein in breast tumor development is mediated by control of MAPK/AP-1 signaling. It is concluded that deactivation of MAPK pathway lead to down-regulation of the AP-1 activation which in turn down regulates the target gene and may be responsible for controlling breast tumor aggressiveness. Thus, it is suggested that MAPKs (ERK, p38 and JNK), MMP-9 and uPAR may be a potential target for anticancer therapy inhibiting tumor vasculature and invasion stimulated by tumor-associated stroma, and regulating the target gene and may be responsible for the beneficial effects of soy protein.

Abstract 5850: p53 gene expression and nitric oxide levels after artemisinin-caffeine treatment in breast, lungs and liver of DMBA-induced tumorigenesis

Abstract Background: With increasing incidence of cancers globally and limited resources in some affected countries, repurposing existing drugs for reducing tumorigenesis is highly important. Artemisinin and caffeine have potent anti-oxidative and anti-cancer properties, although they are therapies for other diseases. When p53 is activated, it repairs DNA damage, prevents tumorigenesis and induces apoptosis whereas nitric oxide is a pro-apoptotic modulator which can upregulate p53 thereby reducing tumorigenesis. We evaluated the biochemical and p53 gene modulatory effects of doses of artemisinin-caffeine combination on breast, lung and liver tumors in rats induced with DMBA. Methods: After due ethical approval, 30 animals were treated with 40mg/kg single dose of 7,12-dimethylbenzene anthracene (DMBA) as a model for DNA damage and induction of carcinogenesis. Five animals each received low dose artemisinin (4mg/kg), low dose caffeine (25mg/kg), low dose combination of Art + Caff (4+25mg/kg), high dose combination of Art + Caff (8+50 mg/kg) or no treatment. All treatment doses were orally administered daily for two weeks post-DMBA treatment, normal control received saline. Nitric oxide levels were assessed and p53 relative gene expression was determined by primer-specific RT-PCR (GAPDH was used as loading control) and amplicons were resolved by gel electrophoresis. Results: DMBA induced lesions in breast, liver, and lung tissues were evident from histology analysis, compared to untreated group. In all 3 tissues, low dose caffeine and combination of Art + Caff significantly reduced p53 gene expression (p &amp;lt; 0.05), but there was significant increase in of p53 expression in low dose Art and high dose Art + Caff groups, which were similar to DMBA group (p&amp;lt;0.05). There was also decrease in nitric oxide levels in treated groups compared to DMBA group. Conclusions: This study shows a dose-dependent anticancer effects of caffeine and artemisinin combination on p53 gene and nitric oxide regulation and hence can mitigate against tumorigenesis. Citation Format: Titilope M. Dokunmu, Sandra C. Opara, Omiete U. Awani. p53 gene expression and nitric oxide levels after artemisinin-caffeine treatment in breast, lungs and liver of DMBA-induced tumorigenesis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 5850.

Chronic stress caused by maternal separation is a possible risk factor for the development of 7,12-dimethyl benzo anthracene-induced breast tumors in rats

Background: Chronic stress has been investigated as a risk factor for breast cancer. Maternal separation (MS) of rats has been used as a chronic stress model that alters certain systemic functions, such as the immune response. Objective: The aim of this study was to determine the possible effect of MS on the development of breast tumors induced by 7,12-dimethyl benzo anthracene (DMBA). Methodology: postnatal day (PND) 1 female Wistar rats were divided into four experimental groups that either were or were not subjected to MS and either received or did not receive DMBA. For MS, PND 1 to 21 pups were separated from their mothers for 360 min/day. On PND 30, carcinomas were induced in mammary glands using DMBA. Body weight was evaluated, and the injected region was palpated. In addition, the mammary glands were subjected to histological examination, and corticosterone levels were determined in all groups. Results: DMBA-induced groups had significantly lower body weight gain compared with the non-DMBA-induced groups. Maternal separation increased the incidence of preneoplastic changes and breast carcinogenesis in DMBA-treated animals compared with control animals. Corticosterone levels were increased in both DMBA-induced and MS groups without interaction. Conclusion: MS is a possible risk factor for DMBA-induced preneoplastic changes and breast tumors in rats.

CCL2 and IL18 expressions may associate with the anti-proliferative effect of noncontact electro capacitive cancer therapy in vivo

Background: Noncontact Electro Capacitive Cancer Therapy (ECCT) is a novel treatment modality in cancer. Chemokine (C-C motif) ligand 2 (CCL2) has a major role in the outgrowth of metastatic breast cancer. Interleukin 18 (IL18) plays a role in macrophage alteration, which leads to excessive angiogenesis. This study aims to elaborate on the association of CCL2, IL18, IL23α, and TNF-α (tumor necrosis factor-alpha) expression with the anti-proliferative effect of ECCT in rat breast tumor tissue. Methods: Low intensity (18 Vpp) and intermediate frequency (150 kHz) alternating current-electric field (AC-EF) between two capacitive electrodes were exposed as external EF to a rat cage. Twenty-four rats were divided into four groups of six replicates. Breast tumor tissues were collected from 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rats. Two groups were non DMBA-induced rats without ECCT exposure (NINT) and with (NIT). The other two groups were DMBA-induced rats without ECCT exposure (INT) and with (IT). Mammary glands and breast tumor tissues were collected from each group and preserved. Hematoxylin-eosin and immunohistochemistry staining were performed on paraffin sections of tissues using anti-PCNA, anti-ErbB2, anti-Caspase3, and anti-CD68. CCL2, IL18, IL23α, and TNF-α mRNA relative expressions were analyzed using qRT-PCR. Results: ECCT exposure may cause the reduction of PCNA protein expression as well as ErbB2 on breast tumor tissues, but it causes the increase of Caspase3 and macrophage CD68 protein. In rat breast tumor tissues of IT groups, the mRNA expression of CCL2 and IL18 are significantly down-regulated, in contrast with the up-regulated expression of these cytokines in tumor tissues of the INT group. IL23α and TNF- α expression remained similar in both groups. Conclusion: CCL2 and IL18 expressions have an association with the inhibition of breast tumor cell proliferation affected by ECCT exposure.

CCL2 and IL18 expressions may associate with the anti-proliferative effect of noncontact electro capacitive cancer therapy in vivo

Background: Noncontact Electro Capacitive Cancer Therapy (ECCT) is a novel treatment modality in cancer. Chemokine (C-C motif) ligand 2 (CCL2) has a major role in the outgrowth of metastatic breast cancer. Interleukin 18 (IL18) plays a role in macrophage alteration, which leads to excessive angiogenesis. This study aims to elaborate on the association of CCL2, IL18, IL23α, and TNF-α (tumor necrosis factor-alpha) expression with the anti-proliferative effect of ECCT in rat breast tumor tissue. Methods: Low intensity (18 Vpp) and intermediate frequency (150 kHz) alternating current-electric field (AC-EF) between two capacitive electrodes were exposed as external EF to a rat cage. Twenty-four rats were divided into four groups of six replicates. Breast tumor tissues were collected from 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rats. Two groups were non DMBA-induced rats without ECCT exposure (NINT) and with (NIT). The other two groups were DMBA-induced rats without ECCT exposure (INT) and with (IT). Mammary glands and breast tumor tissues were collected from each group and preserved. Hematoxylin-eosin and immunohistochemistry staining were performed on paraffin sections of tissues using anti-PCNA, anti-ErbB2, anti-Caspase3, and anti-CD68. CCL2, IL18, IL23α, and TNF-α mRNA relative expressions were analyzed using qRT-PCR. Results: ECCT exposure may cause the reduction of PCNA protein expression as well as ErbB2 on breast tumor tissues, but it causes the increase of Caspase3 and macrophage CD68 protein. In rat breast tumor tissues of IT groups, the mRNA expression of CCL2 and IL18 are significantly down-regulated, in contrast with the up-regulated expression of these cytokines in tumor tissues of the INT group. IL23α and TNF- α expression remained similar in both groups. Conclusion: CCL2 and IL18 expressions have an association with the inhibition of breast tumor cell proliferation affected by ECCT exposure

Mitochondrial dysfunction generates aggregates that resist lysosomal degradation in human breast cancer cells

Disrupting functional protein homeostasis is an established therapeutic strategy for certain tumors. Ongoing studies are evaluating autophagy inhibition for overcoming chemotherapeutic resistance to such therapies by neutralizing lysosomal pH. New and sensitive methods to monitor autophagy in patients are needed to improve trial design and interpretation. We report that mitochondrial-damaged breast cancer cells and rat breast tumors accumulate p53-positive protein aggregates that resist lysosomal degradation. These aggregates were localized to enzymatically-active autolysosomes that were degrading autophagosomes and the autophagic receptor proteins TAX1BP1 and NDP52. NDP52 was identified to associate with aggregated proteins and knocking down NDP52 led to the accumulation of protein aggregates. TAX1BP1 was identified to partly localize with aggregates, and knocking down TAX1BP1 enhanced aggregate formation, suppressed autophagy, impaired NDP52 autophagic degradation and induced cell death. We propose that quantifying aggregates and autophagic receptors are two potential methods to evaluate autophagy and lysosomal degradation, as confirmed using primary human tumor samples. Collectively, this report establishes protein aggregates and autophagy receptors, TAX1BP1 and NDP52, as potential endpoints for monitoring autophagy during drug development and clinical studies.

Antisecretory factor AF-16 improves vascular access to a rat mammary tumour.

Tumor tissue often has an insufficient nutritional supply, in part due to compression of the vascular network from an increased interstitial fluid pressure. We have shown that the antisecretory factor peptide AF-16 can reduce this pressure in experimental rat breast tumors. In this work we studied if AF-16 administration opened up to an increased vascular volume in these tumors. Sprague-Dawley rats were given dimethylbenxanthracene and developed mammary tumors which were studied. Evans Blue was used as an intravascular volume indicator. Under anesthesia the rats were given AF-16 or solvent intranasally, and Evans Blue was injected i.v. 45 min later. Tumors and various organs were dissected and Evans Blue was extracted and colorimetrically quantified. Tumors had a significantly higher vascular volume after AF-16 administration as compared to other organs. Liver and renal vascular volumes were also increased but to a lesser degree than in the tumors. The results indicate that AF16 could be a candidate for increasing vascular access for chemotherapy in cancer therapy.

Local and Systemic Antitumor Effects of Photo-activatable Paclitaxel Prodrug on Rat Breast Tumor Models.

We demonstrated that a large primary and a small untreated distant breast cancer could be controlled by local treatment with our light-activatable paclitaxel (PTX) prodrug. We hypothesized that the treated tumor would be damaged by the combinational effects of photodynamic therapy (PDT) and locally released PTX and that the distant tumor would be suppressed by systemic antitumor effects. Syngeneic rat breast cancer models (single- and two-tumor models) were established on Fischer 344 rats by subcutaneous injection of MAT B III cells. The rats were injected with PTX prodrug (dose: 1umolekg-1 , i.v.), and tumors were treated with illumination using a 690-nm laser (75 or 140mWcm-1 for 30min, cylindrical light diffuser, drug-light interval [DLI] 9h). Larger tumors (~16mm) were effectively ablated (100%) without recurrence for >90days. All cured rats rejected rechallenged tumor for up to 12months. In the two-tumor model, the treatment of the local large tumor (~16mm) also cured the untreated tumor (4-6mm) through adaptive immune activation. This is our first demonstration that local treatment with our PTX prodrug produces systemic antitumor effects. Further investigations are warranted to understand mechanisms and optimal conditions to achieve clinically translatable systemic antitumor effects.

Anticancer activity of Astragalus ovinus against 7, 12 dimethyl benz (a) anthracene (DMBA)-induced breast cancer in rats.

Some species of Astragalus are used for the treatment of various types of cancer. The present study was designed to evaluate the anticancer potential of Astragalus ovinus extract (AOE) against DMBA-induced breast carcinoma in rats. The anti-tumor and antioxidant effects of AOE were evaluated against DMBA-induced breast carcinoma in rats using DPPH, FRAP and ABTS technique, respectively. Forty adult female Sprague-Dawley rats were randomly divided into four groups including the control group received a single dose of DMBA solvent orally, and groups II, III and IV received a single dose of DMBA (40 mg/kg) dissolved in olive oil. Groups I and II received normal saline and groups III and IV were treated with AOE orally (120 and 240 mg/kg respectively) for 60 consecutive days. Chemopreventive effects were assessed in terms of diameter and volume of tumors, expression levels of PCNA, and serum levels of CA15.3, p53, MDA, CAT, and calcium, and histopathological features. AOE contained a noticeable amount of phenolic and flavonoids compounds. This extract showed a potent antioxidant activity both in vitro and in vivo. AOE significantly decreased the diameter and volume of tumors (p<0.01) and reduced the serum levels of CA15.3 (p<0.001), p53 (p<0.01), MDA (p<0.001), and calcium (p<0.01). AOE also decreased the expression of PCNA in cancerous tissues and reduced the histopathological deformity. According to the data, AOE produced a significant chemopreventive activity in DMBA-induced breast tumors in rats, probably due to its antioxidant and its inhibitory effect on some tumorigenicity markers such as CA15.3, p53 and PCNA activity.

CCL2 and IL18 expressions may associate with the anti-proliferative effect of noncontact electro capacitive cancer therapy in vivo.

Background: Noncontact Electro Capacitive Cancer Therapy (ECCT) is a novel treatment modality in cancer. Chemokine (C-C motif) ligand 2 (CCL2) has a major role in the outgrowth of metastatic breast cancer. Interleukin 18 (IL18) plays a role in macrophage alteration, which leads to excessive angiogenesis. This study aims to elaborate on the association of CCL2, IL18, IL23α, and TNF-α (tumor necrosis factor-alpha) expression with the anti-proliferative effect of ECCT in rat breast tumor tissue. Methods: Low intensity (18 Vpp) and intermediate frequency (150 kHz) alternating current-electric field (AC-EF) between two capacitive electrodes were exposed as external EF to a rat cage. Twenty-four rats were divided into four groups of six replicates. Breast tumor tissues were collected from 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rats. Two groups were none DMBA-induced rats without ECCT exposure (NINT) and with (NIT). The other two groups were DMBA-induced rats without ECCT exposure (INT) and with (IT). Mammary glands and breast tumor tissues were collected from each group and preserved. Hematoxylin-eosin and immunohistochemistry staining were performed on paraffin sections of tissues using anti-PCNA, anti-ErbB2, anti-Caspase3, and anti-CD68. CCL2, IL18, IL23α, and TNF-α mRNA relative expressions were analyzed using qRT-PCR. Results: ECCT exposure may cause the reduction of PCNA protein expression as well as ErbB2 on breast tumor tissues, but it causes the increase of Caspase3 and macrophage CD68 protein. In rat breast tumor tissues of IT groups, the mRNA expression of CCL2 and IL18 are significantly down-regulated, in contrast with the up-regulated expression of these cytokines in tumor tissues of the INT group. IL23α and TNF- α expression remained similar in both groups. Conclusion: CCL2 and IL18 expressions have an association with the inhibition of breast tumor cell proliferation affected by ECCT exposure.

Oxygen‐sensitive MRI assessment of tumor response to hypoxic gas breathing challenge

Oxygen-sensitive MRI has been extensively used to investigate tumor oxygenation based on the response (R2 * and/or R1 ) to a gas breathing challenge. Most studies have reported response to hyperoxic gas indicating potential biomarkers of hypoxia. Few studies have examined hypoxic gas breathing and we have now evaluated acute dynamic changes in rat breast tumors. Rats bearing syngeneic subcutaneous (n=15) or orthotopic (n=7) 13762NF breast tumors were exposed to a 16% O2 gas breathing challenge and monitored using blood oxygen level dependent (BOLD) R2 *and tissue oxygen level dependent (TOLD) T1 -weighted measurements at 4.7T. As a control, we used a traditional hyperoxic gas breathing challenge with 100% O2 on a subset of the subcutaneous tumor bearing rats (n=6). Tumor subregions identified as responsive on the basis of R2 * dynamics coincided with the viable tumor area as judged by subsequent H&E staining. As expected, R2 * decreased and T1 -weighted signal increased in response to 100% O2 breathing challenge. Meanwhile, 16% O2 breathing elicited an increase in R2 *, but divergent response (increase or decrease) in T1 -weighted signal. The T1 -weighted signal increase may signify a dominating BOLD effect triggered by 16% O2 in the relatively more hypoxic tumors, whereby the influence of increased paramagnetic deoxyhemoglobin outweighs decreased pO2 . The results emphasize the importance of combined BOLD and TOLD measurements for the correct interpretation of tumor oxygenation properties.

Breast tumor hemodynamic response during a breath-hold as a biomarker to predict chemotherapeutic efficacy: preclinical study.

Continuous wave diffuse optical tomographic/spectroscopic system does not provide absolute concentrations of chromophores in tissue and monitor only the changes of chromophore concentration. Therefore, it requires a perturbation of physiological signals, such as blood flow and oxygenation. In that sense, a few groups reported that monitoring a relative hemodynamic change during a breast tissue compression or a breath-hold to a patient can provide good contrast between tumor and nontumor. However, no longitudinal study reports the utilization of a breath-hold to predict tumor response during chemotherapy. A continuous wave near-infrared spectroscopy was employed to monitor hemodynamics in rat breast tumor during a hyperoxic to normoxic inhalational gas intervention to mimic a breath-hold during tumor growth and chemotherapy. The reduced oxyhemoglobin concentration during inhalational gas intervention correlated well with tumor growth, and it responded one day earlier than the change of tumor volume after chemotherapy. In conclusion, monitoring tumor hemodynamics during a breath-hold may serve as a biomarker to predict chemotherapeutic efficacy of tumor.

The combined therapeutic effects of 131iodine-labeled multifunctional copper sulfide-loaded microspheres in treating breast cancer

Compared to conventional cancer treatment, combination therapy based on well-designed nanoscale platforms may offer an opportunity to eliminate tumors and reduce recurrence and metastasis. In this study, we prepared multifunctional microspheres loading 131I-labeled hollow copper sulfide nanoparticles and paclitaxel (131I-HCuSNPs-MS-PTX) for imaging and therapeutics of W256/B breast tumors in rats. 18F-fluordeoxyglucose (18F-FDG) positron emission tomography/computed tomography (PET/CT) imaging detected that the expansion of the tumor volume was delayed (P<0.05) following intra-tumoral (i.t.) injection with 131I-HCuSNPs-MS-PTX plus near-infrared (NIR) irradiation. The immunohistochemical analysis further confirmed the anti-tumor effect. The single photon emission computed tomography (SPECT)/photoacoustic imaging mediated by 131I-HCuSNPs-MS-PTX demonstrated that microspheres were mainly distributed in the tumors with a relatively low distribution in other organs. Our results revealed that 131I-HCuSNPs-MS-PTX offered combined photothermal, chemo- and radio-therapies, eliminating tumors at a relatively low dose, as well as allowing SPECT/CT and photoacoustic imaging monitoring of distribution of the injected agents non-invasively. The copper sulfide-loaded microspheres, 131I-HCuSNPs-MS-PTX, can serve as a versatile theranostic agent in an orthotopic breast cancer model.

In vivo Raman spectroscopy of breast tumors prephotodynamic and postphotodynamic therapy

AbstractBreast cancer is most fatal cancer among women worldwide. The high mortality can be attributed to late detection and low treatment efficacy. Treatment is difficult owing to the multitude of breast cancer subtypes and making decisions on therapeutic strategy difficult. A tool to predict treatment prognosis may greatly aid this decision making. Currently available prediction methods have low accuracy in addition to several other disadvantages. Of the several new techniques being investigated for prognosis prediction, Raman spectroscopy (RS) has the advantage of high sensitivity, rapidity, and amenability to in vivo applications, making it ideal for clinical translation. In this study, we have evaluated the biochemical changes posttherapy with respect to pretherapy using RS. In vivo Raman spectra acquired from live rat breast tumors (skin removed) without treatment, 24 hr postphotosensitizer injection and 24 hr after photodynamic therapy, were analyzed using multivariate principal component—linear discriminant analysis. Relative increase in some spectral signatures associated with nucleic acids, amino acids, and proteins, especially collagen, were observed posttherapy; and pretherapy and posttherapy spectra could be classified with 100% efficiency. The ability of RS to detect these changes suggests possibility of deciphering spectral markers for prognostic applications in future.

Hyperthermia therapy monitoring with guidance of B-Mode ultrasound: Study on rat breast tumor tissue

Local hyperthermia treatment for cáncer therapy has shown increasing interest in conjunction to the other existence treatment as such chemotherapy, clinical surgery and radiation. B-Mode ultrasound have shown great potential for local hyperthermia, as it is nonionizing, convenient, and has relatively simple signal processing and it offers good spatial resolution.The aim of this study is to compare the most optimum B-Mode ultrasound parameters in normal and pathological tissue. The subjects were dissected and exposed to hyperthermia at 37◦C, 40◦C, 45◦C, 50◦C and 55◦C. Result shows, for mean grey scale in normal and pathological tissue, 37◦C and 40◦C was optimum temperature with correspond values of 188.08 and 199.26 meanwhile for pixel deviation analysis, at 45◦C and 55◦C the best temperature is observed in both tissue with pixel deviation of 57.27 and 57.25. Additionally, it is proven with accuracy of testing to validation data using Artificial Neural Network, ANN, about 88.89% and 81.25% was achieved.