Effect of preoperative FOLFOX chemotherapy on CCL20/CCR6 expression in colorectal liver metastases

AIM： To evaluate the influence of preoperative FOLFOX chemotherapy on CCL20/CCR6 expression in liver metastases of stage IV colorectal cancer （CRC） patients.（/7 = 53） and in patients who did not receive FOLFOX chemotherapy prior to liver surgery （n = 29）. RESULTS： Of the 53 patients who received FOLFOX, time to liver surgery was n〈 1 mo in 14 patients, 〈 1 year in 22 patients and 〉 1 year in 17 patients, respectively. In addition, we investigated the proliferation rate of CRC cells in liver metastases in the different patient groups. Both CCL20 and CCR6 mRNA and protein ex- pression levels were significantly increased in patients who received preoperative FOLFOX chemotherapy ~〈 12 mo before liver surgery （P 〈 0.001） in comparison to patients who did not undergo FOLFOX treatment. Further, proliferation of CRLM cells as measured by Ki-67 was increased in patients who underwent FOLFOX treat- ment. CCL20 and CCR6 expression levels were significantly increased in CRLM patients who had undergone preoperative FOLFOX chemotherapy. CONCLUSION： This chemokine/receptor up-regulation could lead to increased proliferation/migration through an autocrine mechanism which might be used by surviving metastatic cells to escape cell death caused by FOLFOX.

High mobility group-box 3 overexpression is associated with poor prognosis of resected gastric adenocarcinoma

AIM:To elucidate high mobility group-box 3(HMGB3) protein expression in gastric adenocarcinoma,its potential prognostic relevance,and possible mechanism of action.METHODS:Ninety-two patients with gastric adenocarcinomas surgically removed entered the study.HMGB3 expression was determined by immunohistochemistry through a tissue microarray procedure.The clinicopathologic characteristics of all patients were recorded,and regular follow-up was made for all patients.The inter-relationship of HMGB3 expression with histological and clinical factors was analyzed using nonparametric tests.Survival analysis was carried out by Kaplan-Meier(log-rank) and multivariate Cox(Forward LR) analyses between the group with overexpression of HMGB3 and the group with low or no HMGB3 ex-pression to determine the prognosis value of HMGB3 expression on overall survival.Further,HMGB3 expression was knocked down by small hairpin RNAs(shRNAs) in the human gastric cancer cell line BGC823 to observe its influence on cell biological characteristics.The MTT method was utilized to detect gastric cancer cell proliferation changes,and cell cycle distribution was analyzed by flow cytometry.RESULTS:Among 92 patients with gastric adenocarcinomas surgically removed in this study,high HMGB3 protein expression was detected in the gastric adenocarcinoma tissues vs peritumoral tissues(P < 0.001).Further correlation analysis with patients' clinical and histology variables revealed that HMGB3 overexpression was obviously associated with extensive wall penetration(P = 0.005),a positive nodal status(P = 0.004),and advanced tumor-node-metastasis(TNM) stage(P = 0.001).But there was no correlation between HMGB3 overexpression and the age and gender of the patient,tumor localization or histologic grade.Statistical Kaplan-Meier survival analysis disclosed significant differences in overall survival between the HMGB3 overexpression group and the HMGB3 no or low expression group(P = 0.006).The expected overall survival time was 31.00 ± 3.773 mo(95%CI = 23.605-38.395) for patients with HMGB3 overexpression and 49.074 ± 3.648 mo(95%CI = 41.925-57.311) for patients with HMGB3 no and low-level expression.Additionally,older age(P = 0.040),extensive wall penetration(P = 0.008),positive lymph node metastasis(P = 0.005),and advanced TNM tumor stage(P = 0.007) showed negative correlation with overall survival.Multivariate Cox regression analysis indicated that HMGB3 overexpression was an independent variable with respect to age,gender,histologic grade,extent of wall penetration,lymph nodal metastasis,and TNM stage for patients with resectable gastric adenocarcinomas with poor prognosis(hazard ratio = 2.791,95%CI = 1.233-6.319,P = 0.019).In the gene function study,after HMGB3 was knocked down in the gastric cell line BGC823 by shRNA,the cell proliferation rate was reduced at 24 h,48 h and 72 h.Compared to BGC823 shRNA-negative control(NC) cells,the cell proliferation rate in cells that had HMGB3 shRNA transfected was significantly decreased(P < 0.01).Finally,cell cycle analysis by FACS showed that BGC823 cells that had HMGB3 knocked down were blocked in G1/G0 phase.The percentage of cells in G1/G0 phase in BGC823 cells with shRNA-NC and with shRNA-HMGB3 was 46.84% ± 1.7%,and 73.03% ± 3.51% respectively(P = 0.001),whereas G2/M cells percentage decreased from 26.51% ± 0.83% to 17.8% ± 2.26%.CONCLUSION:HMGB3 is likely to be a useful prognostic marker involved in gastric cancer disease onset and progression by regulating the cell cycle.

The Inhibitory Effect of Endostatin and Doxycycline Administration on B16 Melanoma Angiogenesis and Cellular Proliferation

OBJECTIVE To investigate the effect of endostatin and doxycycline on melanoma cellular proliferation and tumor angiogenesis.METHODS The effects of endostatin and doxycycline were studied in mice transplanted with B16 melanoma cells. The mice were divided into 4 groups that were treated as follows： endostatin treatment （E group）, doxycycline treatment （D group）, endostatin plus doxycycline trearment （DE group）, controls （C group） received no treatment. Following 9 days of treatment the tumor tissue was removed to compare the differences in the tumor necrotic rate and micro-vessel density （MVD） among the different groups. Immunohistochemical staining was conducted to detect the expression of proliferating cell nuclear antigen （PCNA） in the different groups.RESULTS The MVD of the 3 experimental groups was significantly less than the control group, （F = 10.888, P 〈 0.05）, indicating that doxycycline and endostatin can inhibit tumor angiogenesis by decreasing the tumor blood supply. This effect results in inhibition of tumor cellular proliferation and promotion of tumor cell necrosis. The tumor cell necrotic rate of the 3 experimental groups were all significantly higher than the C group （F = 7.229, P 〈 0.05） and the difference between the DE and C groups also was statistically significant. PCNA expression in all 3 experimental groups was statistically less than the C group （F = 17.729, P 〈 0.05）.CONCLUSION The combined use of endostatin and doxycycline in vivo can influence PCNA expression and angiogenesis in melanoma, and significantly inhibit melanoma cellular proliferation.

Adenovirus-mediated PTEN gene transfection suppresses growth and promotes chemosensitivity of endometrial carcinoma

Objective： To determine the potential of sustained transgene expression by intratumoral injection of Ad-PTEN in the nude mouse model of endometrial carcinoma. Methods and Results： We constructed recombinant adenovirus carrying the wild-type PTEN gene （Ad-PTEN）. RL95-2 cells, an endometrial carcinoma cell line lacking PTEN function, was infected with Ad-PTEN and showed increased expression of PTEN and chemosensitivity to doxorubicin, decreased proliferation rate, and elevated apoptosis and Go/G1 arrest. Furthermore, the tumorigenicity of these cells was also completely suppressed. These results indicated that gene therapy with Ad-PTEN could significantly inhibit the endometrial carcinoma xenografts growth in nude mice by intratumoral injection, induce apoptosis of tumor cells, and reduce expression of proliferating cell nuclear antigen （PCNA）. Immunohistochemistry analysis also showed that the expression of progesterone receptors （PR） in Ad-PTEN treated tumor cells were induced, while P-glycoproteins （P-gp） and estrogen receptors （ER） decreased significantly. Conclusion： PTEN may play an important role in the development of endometrial carcinoma. Our findings cast new lights for treatment ofendometrial carcinoma.

In vitro incubation of cytokine-induced killer cells from patients with and without hepatitis B virus and a cell subset analysis

Objective The aim of the study was to explore the difference between immune cell subsets during the incubation of cytokine-induced kill cells （CIKs） from patients with and without hepatitis B virus （HBV）. Methods Peripheral blood samples were extracted from 50 tumor patients, and were divided into two groups according to the presence or absence of HBV. The proliferation rate and activity of CIK cells were examined based on counts on days 1, 5, 7, 9, 11, 13, and 15 of culture. Additionally, the CD3＋, CD4＋, CD8＋, CD3＋CD8＋, C＋）3＋CD4＋, and CD3＋CD56＋ T cell populations were analyzed by flow cytometry on days 5, 7, 10, 13, and 15 of culture. Results Proliferation over a 15-day period was higher in the HBV-positive group than in the negative group （280-fold vs. 180-fold increase, respectively）, but there was no significant difference between the two groups at each time point. The frequencies of CD3＋, CD8＋ T, CD3＋CD8＋, and CD3＋CD56＋T cells increased over time, while those of CD4＋ and CD3＋CD4＋ T cells decreased over time, and these changes were greater in the positive group than in the negative group. The differences in CD8＋ T cells and CD3＋CD4＋ T cells between the two groups were significant （P 〈 0.05）. Conclusion The proliferative capacity of CIK cells was higher for patients in the HBV-positive group than those in the HBV-negative group, and immune cell subsets were more favorable in the HBV-positive group than the neaative arouD.

Effect of Abscisic Acid on NaCI Stressed Callus Proliferation and Plant Regeneration in Rice

In rice, 2,4-Dichlorphenoxyacetic acid （MS2） has been considered best for callus induction and combination ofNAA, IAA, ABA （MS3） for somatic embryogenesis. Efficient plant regeneration was observed when MS basal salts supplied with 3% sorbitol and 3% maltose as carbon sources without hormones （MSac）. During this experiment, effect of sodium chloride （NaCl） and abscisic acid （ABA） was assessed on callus growth, plant regeneration and root induction efficiency of rice （Oryza sativa L.） varieties IR-6 and Basmati-370. Callus proliferation rate was highly decreased in both varieties on MS2b （100 mol.m3 NaCl ＋ 0.5 mg＇L1 ABA） than MS2a （100 mol＇m3 NaCl） cultures significantly. Proline, glycine-betaine and reducing sugars were increased significantly in MS2a and MS2b callusing cultures. However, total proteins were decreased in MS2a, while slightly increased in MS2b. Maximum plant regeneration （9.42 ±0.54 and 10.67 ±0.50 plantlets.callus1） from somatic embryos was observed on MS4c in IR-6 and Basmati-370, while 1.56 ± 0.06 （IR-6） and 0.95 ±0.05 （Basmati-370） plantlets callus1 were observed on MSab （100 mol·m3 NaCl）. No plant regeneration was observed on MS4b （100 mol·m3 NaCl ＋ 0.5 mg·L-1 ABA） medium in both varieties. Inhibition of root induction efficiency was high in MSsb （100 mol·m3 NaCl ＋ 0.5 mg·L-1 ABA） than MS5a （100 mol·m3 NaCl） in the stressed cultures （P 〈 0.05）. In this experiment, it was concluded that ABA involved in somatic embryogenesis and elevation of NaCl stress, while it causes inhibition of cell＇s growth as well as its regeneration into plantlets from somatic embryos.

Cell cycle regulator geminin is dispensable for the proliferation of vascular smooth muscle cells

The proliferation of vascular smooth muscle cells(VSMCs) plays a major role in the pathogenesis of many cardiovascular diseases.Geminin regulates DNA replication and cell cycle progression and plays a key role in the proliferation of cancer cells.We therefore hypothesized that geminin regulates the proliferation of VSMCs.The present study demonstrates that the level of geminin expression was low in quiescent VSMCs(approximately 90% and 10% of cells in the G1 and in S/G2/M phases of the cell cycle,respectively),increased as more cells entered in S/G2/M,and then decreased as cells exited S/G2/M.Further,angiotensin II and norepinephrine stimulated expression of geminin in VSMCs.However,the DNA content,nuclear morphology,percentage of cells at different stages of the cell cycle,and rate of proliferation of VSMCs from which geminin was either depleted or overexpressed were all similar.These findings indicate geminin functions differently in VSMCs than it does in cancer cell lines and that it may provide a target for treating cancers without affecting normal cells.

HOW SELF-PROLIFERATION OF CD4＋T CELLS AFFECT THE HIV DYNAMICS IN AN IN-HOST TARGET-CELL LIMITED HIV MODEL WITH SATURATION INFECTION RATE： A QUASI-STEADY-STATE APPROXIMATION ANALYSIS

In this study, we consider two target-cell limited models with saturation type infec- tion rate and intracellular delay： one without self-proliferation and the other with self- proliferation of activated CD4＋T cells. We discuss about the local and global behavior of both the systems in presence and absence of intracellular delay. It is shown that the endemic equilibrium of a target-cell limited model would be unstable in presence and absence of intraeellular delay only when self-proliferation of activated CD4＋T cell is considered. Otherwise, all positive solutions converge to the endemic equilibrium or disease-free equilibrium depending on whether the basic reproduction ratio is greater than or less than unity. Our study suggests that amplitude of oscillation is negatively correlated with the constant input rate of CD4＋T cell when intracellular delay is absent or low. However, they are positively correlated if the delay is too high. Amplitude of oscillation, on the other hand, is always positively correlated with the proliferation rate of CD4＋T cell for all delay. Our mathematical and simulation analysis also suggest that there are many potential contributors who are responsible for the variation of CD4＋T cells and virus particles in the blood plasma of HIV patients.