Wenhong Xu

Preoperatively molecular staging with CM10 ProteinChip and SELDI-TOF-MS for colorectal cancer patients

Objectives: To detect the serum proteomic patterns by using SELDI-TOF-MS (surface enhanced laser desorption/ionization-time of flight-mass spectrometry) technology and CM10 ProteinChip in colorectal cancer (CRC) patients, and to evaluate the significance of the proteomic patterns in the tumour staging of colorectal cancer. Methods: SELDI-TOF-MS and CM10 ProteinChip were used to detect the serum proteomic patterns of 76 patients with colorectal cancer, among them, 10 Stage I, 19 Stage II, 16 Stage III and 31 Stage IV samples. Different stage models were developed and validated by support vector machines, discriminant analysis and time-sequence analysis. Results: The Model I formed by 6 protein peaks (m/z: 2759.58, 2964.66, 2048.01, 4795.90, 4139.77 and 37761.60) could be used to distinguish local CRC patients (Stage I and Stage II) from regional CRC patients (Stage III) with an accuracy of 86.67% (39/45). The Model II formed by 3 protein peaks (m/z: 6885.30, 2058.32 and 8567.75) could be used to distinguish locoregional CRC patients (Stage I, Stage II and Stage III) from systematic CRC patients (Stage IV) with an accuracy of 75.00% (57/76). The Model III could distinguish Stage I from Stage II with an accuracy of 86.21% (25/29). The Model IV could distinguish Stage I from Stage III with accuracy of 84.62% (22/26). The Model V could distinguish Stage II from Stage III with accuracy of 85.71% (30/35). The Model VI could distinguish Stage II from Stage IV with accuracy of 80.00% (40/50). The Model VII could distinguish Stage III from Stage IV with accuracy of 78.72% (37/47). Different stage groups could be distinguished by the two-dimensional scattered spots figure obviously. Conclusion: This method showed great success in preoperatively determining the colorectal cancer stage of patients.

Doxorubicin-mediated radiosensitivity in multicellular spheroids from a lung cancer cell line is enhanced by composite micelle encapsulation

Background The purpose of this study is to evaluate the efficacy of composite doxorubicinloaded micelles for enhancing doxorubicin radiosensitivity in multicellular spheroids from a non-small cell lung cancer cell line. Methods A novel composite doxorubicin-loaded micelle consisting of polyethylene glycolpolycaprolactone/Pluronic P105 was developed, and carrier-mediated doxorubicin accumulation and release from multicellular spheroids was evaluated. We used confocal laser scanning microscopy and flow cytometry to study the accumulation and efflux of doxorubicin from A549 multicellular spheroids. Doxorubicin radiosensitization and the combined effects of irradiation and doxorubicin on cell migration and proliferation were compared for the different doxorubicin delivery systems. Results Confocal laser scanning microscopy and quantitative flow cytometry studies both verified that, for equivalent doxorubicin concentrations, composite doxorubicin-loaded micelles significantly enhanced cellular doxorubicin accumulation and inhibited doxorubicin release. Colony-forming assays demonstrated that composite doxorubicin-loaded micelles are radiosensitive, as shown by significantly reduced survival of cells treated by radiation + composite micelles compared with those treated with radiation + free doxorubicin or radiation alone. The multicellular spheroid migration area and growth ability verified higher radiosensitivity for the composite micelles loaded with doxorubicin than for free doxorubicin. Conclusion Our composite doxorubicin-loaded micelle was demonstrated to have radiosensitization. Doxorubicin loading in the composite micelles significantly increased its cellular uptake, improved drug retention, and enhanced its antitumor effect relative to free doxorubicin, thereby providing a novel approach for treatment of cancer.

The crosstalk: Tumor-infiltrating lymphocytes rich in regulatory T cells suppressed cancer-associated fibroblasts

Abstract Background. The interactions between cancer-associated fibroblasts (CAFs) and cancer cells or tumor-infiltrating lymphocytes (TILs) and cancer cells play important roles in cancer progression and metastasis. However, studies related to the crosstalk between CAFs and TILs in tumor microenvironment (TME) are still lacking. In this study, we mainly investigated the interactions between CAFs and TILs. Material and methods. The distribution of TILs rich in regulatory T cells (Tregs) in breast cancer tissues was evaluated using hematoxylin-eosin staining and immunohistochemistry with anti-CD3, anti-Foxp3, and anti-α-smooth muscle actin antibodies. Homologous CAFs/normal fibroblasts (NFs) and TILs cultured in vitro were identified and detected using immunocytochemistry and flow cytometry (FCM). The direct interaction among these cell types was studied via a factorial design in a co-cultured system. Their indirect interaction was assayed using Transwell plates. The cell cycle and apoptosis of CAFs/NFs co-cultured with TILs was analyzed using propidium iodide staining. Results. Histochemistry demonstrated most of the TILs including Tregs, were distributed in the cancer stroma, adjoining to CAFs. This finding implies that both cell types interact closely in the TME. Identification of the cultured cells showed that CAFs maintained their activated phenotype within limited passages in vitro, and that the TILs population contained a high percentage of Tregs. Data analysis of the factorial design suggests significant interactions among CAFs, NFs, and TILs in both direct and indirect contact ways. The CAFs and NFs were suppressed signally by TILs, which are probably induced by the secretory cytokines derived from TILs or Tregs. Although apoptosis was not detected in CAFs/NFs, the cell cycle assay suggested that the CAFs/NFs were arrested in the G2/M phase by the TILs and their secretory cytokines. Conclusion. CAFs and NFs were dramatically suppressed by Tregs-rich TILs. This suggests the interaction between TILs and CAFs might modify the TME in an unknown manner.

Cytohesins/ARNO: The Function in Colorectal Cancer Cells

Epidermal growth factor (EGF) and insulin-like growth factor-I (IGF-I) are critical regulators of cell differentiation, survival, proliferation, and migration in cancers. This study found that ARNO (cytohesin-2), an activator of the EGF and IGF-I pathways, was more highly expressed in colorectal cancer tissue than in benign adjacent colorectal tissue. When ARNO-siRNA or the chemical inhibitor SecinH3 blocked ARNO, the downstream of the EGF and IGF-I pathways decreased in colorectal cell lines HT29 and HCT116. This blocking also weakened cell proliferation, invasion, and migration in vitro. Furthermore, EGF receptor (EGFR)-dependent colorectal tumor xenografts in nude mouse exerted anti-proliferative and growth suppression effects by injecting secineH3. These data suggested that inhibiting cytohesins or ARNO as cytoplasmic activators of EGFR and IGF-I in colorectal cancer resulted in anti-proliferation, reduced invasion, decreased migration, and suppressed growth in vivo and in vitro. Therefore, cytohesins or ARNO may be a potential therapy target for some colorectal cancer.

Levonorgestrel-releasing intrauterine device is an efficacious contraceptive for women with leiomyoma.

Objective: To evaluate the efficacy of a levonorgestrel-releasing intrauterine device (LNG-IUD) as contraception for women with uterine leiomyoma. Methods: Thirty women with uterine leiomyoma requiring contraception were enrolled and had a LNG-IUD inserted. Menstrual blood volume (assessed by a pictorial blood loss assessment chart [PBAC]), haemoglobin concentration, volume of uterus and leiomyoma were determined before and after LNG-IUD insertion. Results: The median (range) PBAC score was 145.0 (60.0 - 232.0) before LNG-IUD insertion, which significantly decreased to 44.0 (0.0 - 99.0) and 36.0 (0.0 - 90.0) after 6 and 12 months of LNG-IUD use, respectively. Prior to LNG-IUD insertion, the median haemoglobin concentration in patients with anaemia was 100.0 (69.0 - 109.0) g/l and this increased significantly after 6 and 12 months of LNG-IUD use, to 117.5 (101.0 - 131.0) g/l and 119.5 (108.0 - 135.0) g/l, respectively. There were no significant changes in uterine or leiomyoma volumes. No patient became pregnant and there were no severe side-effects; one IUD was expulsed. Conclusion: The LNG-IUD may be a safe and effective contraceptive for women of reproductive age with uterine leiomyoma.

Mitochondrial Targeted Doxorubicin-Triphenylphosphonium Delivered by Hyaluronic Acid Modified and pH Responsive Nanocarriers to Breast Tumor: in Vitro and in Vivo Studies

Multidrug resistance (MDR) is the major obstacle for chemotherapy. In a previous study, we have successfully synthesized a novel doxorubicin (DOX) derivative modified by triphenylphosphonium (TPP) to realize mitochondrial delivery of DOX and showed the potential of this compound to overcome DOX resistance in MDA-MB-435/DOX cells. (1) To introduce specificity for DOX-TPP to cancer cells, here we report on the conjugation of DOX-TPP to hyaluronic acid (HA) by hydrazone bond with adipic acid dihydrazide (ADH) as the acid-responsive linker, producing HA- hydra-DOX-TPP nanoparticles. Hyaluronic acid (HA) is a natural water-soluble linear glycosaminoglycan, which was hypothesized to increase the accumulation of nanoparticles containing DOX-TPP in the mitochondria of tumor cells upon systemic administration, overcoming DOX resistance, in vivo. Our results showed HA- hydra-DOX-TPP to self-assemble to core/shell nanoparticles of good dispersibility and effective release of DOX-TPP from the HA- hydra-DOX-TPP conjugate in cancer cells, which was followed by enhanced DOX mitochondria accumulation. The HA- hydra-DOX-TPP nanoparticles also showed improved anticancer effects, better tumor cell apoptosis, and better safety profile compared to free DOX in MCF-7/ADR bearing mice.

Targeting tumor hypoxia with stimulus-responsive nanocarriers in overcoming drug resistance and monitoring anticancer efficacy

Although existing nanomedicines have focused on the tumor microenvironment with the goal of improving the effectiveness of conventional chemotherapy, the penetration of a tumors core still represents a formidable barrier for existing drug delivery systems. Therefore, a novel multifunctional hypoxia-induced size-shrinkable nanoparticle has been designed to increase the penetration of drugs, nucleic acids, or probes into tumors. This cooperative strategy relies on three aspects: (i) the responsiveness of nanoparticles to hypoxia, which shrink when triggered by low oxygen concentrations; (ii) the core of a nanoparticle involves an internal cavity and strong positive charges on the surface to deliver both doxorubicin and siRNA; and (iii) a reactive oxygen species (ROS) probe is incorporated in the nanoparticle to monitor its preliminary therapeutic response in real time, which is expected to realize the enhanced efficacy together with the ability to self-monitor the anticancer activity. A more effective inhibition of tumor growth was observed in tumor-bearing zebrafish, demonstrating the feasibility of this cooperative strategy for in vivo applications. This research highlights a promising value in delivering drugs, nucleic acids, or probes to a tumors core for cancer imaging and treatment. STATEMENT OF SIGNIFICANCE Hypoxia-induced chemoresistance of tumor cells still represents a formidable barrier, as it is difficult for existing drug delivery systems to penetrate the tumor hypoxia core. This study involves the hypoxia-responsive size-shrinkable nanoparticle co-delivery of DOX and siRNA to enhance the penetration of DOX deep within tumors and subsequently disturb crucial pathways of cancer development induced by hypoxia and to improve sensitization to DOX chemotherapy. Furthermore, the nanopreparation can combine the ROS probe as a self-reporting nanopreparation to realize the function of real-time feedback efficacy, which has a good application prospect in the diagnosis and treatment of cancer.

Correction: Cytohesins/ARNO: The Function in Colorectal Cancer Cells

In the Funding section, the grant number from the funder Zhejiang Provincial Natural Science Foundation of China is listed incorrectly. The correct grant number is: LQ13H160015.

Translational medicine research on early detection and early diagnosis of colorectal cancer in China

Materials and methods From 2007, a CRC screening program was begun in three cities (Hangzhou, Haerbing, and Shanghai city, aimed at finding more early CRC cases and premalignant lesions such as adenoma. The two steps screening model was applied. As the primary screening, the Simultaneous iFOBT (immunochemical Fecal Occult Blood Testing) and high risk factors questionnaire investigation (HRFQ) will be employed for selection the high risk population from target population. Then the high risk population will be further selected by colonoscopy.

Gastrointestinal metastasis of primary lung cancer: An analysis of 366 cases

The gastrointestinal (GI) tract is not a common site of metastasis in primary lung cancer. The aim of the present study was to reveal the clinical and prognostic characteristics of gastrointestinal metastases of lung cancer (GMLC). Information on 366 cases of GMLC was collected and factors that affect severe GI complications were analyzed. Univariate and multivariate survival analyses were performed using the Cox proportional hazards model. Of the cases analyzed, the small intestine (59.6%) and colorectum (25.6%) were the two organs where lung cancer was most likely to metastasize in the GI tract. Squamous cell carcinoma (28.5%), adenocarcinoma (27.6%) and large cell carcinoma (20.9%) were the three most common histological types. However, compared with the histological distributions of primary lung cancer, patients with large cell carcinoma exhibited the highest elevated risk of GMLC [relative risk (RR), 4.07; P<0.001] and those with adenocarcinoma exhibited the lowest risk (RR, 0.58; P<0.001). Differences in organ involvement and in histological type led to varying GI complications. It was also indicated that chemotherapy was associated with a decreased risk of hemorrhage (P=0.006), but there was no reduction in the risk of hemorrhage associated with perforation and obstruction (P>0.05). The median overall survival time of GMLC patients was 2.8 months (range, 0–108 months). The survival analyses revealed that perforation and extra-GI metastasis were negative prognostic factors but abdominal surgery was identified a positive prognostic factor. In conclusion, the histological distribution of GMLC differed from that of primary lung cancer. Sufficient and careful patient evaluation, targeted surgeries and systemic therapies for specific patients are able to increase patient survival rate and improve the quality of life.