Laboratory indicators predict axillary nodal pathologic complete response after neoadjuvant therapy in breast cancer.

The purpose was to integrate clinicopathological and laboratory indicators to predict axillary nodal pathologic complete response (apCR) after neoadjuvant therapy (NAT). The pretreatment clinicopathological and laboratory indicators of 416 clinical nodal-positive breast cancer patients who underwent surgery after NAT were analyzed from April 2015 to 2020. Predictive factors of apCR were examined by logistic analysis. A nomogram was built according to logistic analysis. Among the 416 patients, 37.3% achieved apCR. Multivariate analysis showed that age, pathological grading, molecular subtype and neutrophil-to-lymphocyte ratio were independent predictors of apCR. A nomogram was established based on these four factors. The area under the curve (AUC) was 0.758 in the training set. The validation set showed good discrimination, with AUC of 0.732. In subtype analysis, apCR was 23.8, 47.1 and 50.8% in hormone receptor-positive/HER2-, HER2+ and triple-negative subgroups, respectively. According to the results of the multivariate analysis, pathological grade and fibrinogen level were independent predictors of apCR after NAT in HER2+ patients. Except for traditional clinicopathological factors, laboratory indicators could also be identified as predictive factors of apCR after NAT. The nomogram integrating pretreatment indicators demonstrated its distinguishing capability, with a high AUC, and could help to guide individualized treatment options.

Lay abstract The purpose of this study was to integrate more pretreatment indicators, including clinicopathological factors and simple laboratory indicators, to predict axillary nodal pathologic complete response (apCR) after neoadjuvant therapy for breast cancer. The authors collected the pretreatment clinicopathological factors and laboratory indexes of 416 nodal-positive patients with breast cancer. The authors then built a nomogram to predict the therapeutic effect in axillary lymph nodes. Among 416 patients, 37.3% (155 of 416) achieved apCR. The results showed that age, pathological grading, molecular subtype and neutrophil-to-lymphocyte ratio were independent predictors of apCR. Based on these four factors, a nomogram was then built. This nomogram helped to predict apCR. In addition to traditional clinicopathological factors, laboratory indicators were also identified as predictive factors of apCR after neoadjuvant therapy. Integrating pretreatment indicators might help to predict apCR and guide individualized treatment options.

Efficacy and safety analysis of paclitaxel, docetaxel and liposomal paclitaxel after neoadjuvant therapy in breast cancer.

BACKGROUND AND PURPOSE: Paclitaxel-based regimens are widely used in the neoadjuvant therapy (NAT) of breast cancer. The purpose is to analysis the efficacy and adverse events (AEs) among common paclitaxel (PTX), docetaxel and liposomal paclitaxel. At the same time, we want to analysis the axillary nodal pathologic complete response (apCR) after NAT among the three groups. METHODS: From April 2014 to 2020, 647 breast cancer patients underwent operation after NAT were included in this study. Patients received full course of anthracycline- and paclitaxel-based chemotherapy before surgery. The paclitaxel-based regimens included PTX, docetaxel and liposomal paclitaxel. The therapy efficacy and AEs of the three groups were evaluated. At the same time, the apCR was also analyzed. RESULTS: In general, 30.6% (198/647) of patients achieved breast pathologic complete response (bpCR), which was 28.6%, 28.3% and 39.3% among PTX, docetaxel and liposomal paclitaxel group, respectively (p = 0.067). The total pathologic complete response (tpCR) (achieving both bpCR and apCR) was 21.6% (140/647). The tpCR was 13.3%, 19.4% and 34.4% among PTX, docetaxel and liposomal paclitaxel group, respectively (p = 0.026). The multivariate logistic analysis result showed that clinical tumor stage and molecular subtype were significantly associated with tpCR (all p < 0.05). Among 592 clinical positive patients (cN+), the apCR was 39.0% (231/592). The multivariate logistic analysis showed that paclitaxel- based regimens and molecular subtype were indicated as independent predictors for apCR of NAT. The apCR was significantly higher in liposomal paclitaxel group (63.5%) than in PTX (24.6%) and docetaxel group (34.8%) (p < 0.001). The subgroup analysis among different molecular subtypes found that in triple negative (TN) and HER-2 positive (HER2+) subgroup, the apCR in liposomal paclitaxel group was significantly higher than those in PTX and docetaxel group (all p < 0.05). But no significant result was found in the subgroup analysis in hormone receptor positive/HER-2 negative subgroup (p = 0.050). Safety analysis indicated that the incidence of neutropenia (grade III-IV) and peripheral neurotoxicity (grade I-II) was significantly lower in the liposomal paclitaxel group than in the PTX and docetaxel group. The incidence of oral mucositis, anaphylaxis and palmar-plantar erythrodysesthesia syndrome was also much lower in liposomal paclitaxel than other two groups (all p < 0.05). And there was no significant difference in other AEs among the three groups (all p > 0.05). CONCLUSION: Liposome paclitaxel had similar tumor suppressor effect compared with PTX and docetaxel in NAT setting. Moreover, it had a better axillary lymph node (ALN) response after NAT than PTX and docetaxel. These patients who received liposome paclitaxel had more chance to avoid ALN dissection after NAT. At the same time, the application of liposome enables liposome paclitaxel could significantly reduce AEs caused by chemotherapy. Therefore, we suggested the application of liposome paclitaxel in the NAT setting, especially for cN+ patients with TN and HER2 + disease.

Twist may be associated with invasion and metastasis of hypoxic NSCLC cells.

Hypoxia promotes tumor invasion and metastasis via multiple mechanisms, including epithelial-mesenchymal transition (EMT). Twist, an EMT regulator, has been disclosed to associate with invasion and metastasis as well as poor prognosis of many malignancies. However, it remains undefined whether Twist is involved in invasion and metastasis of hypoxic non-small cell lung cancer (NSCLC). In this study, protein levels of Twist, hypoxia-inducible factor-1α (HIF-1α), and EMT markers (E-cadherin and vimentin) were examined by immunohistochemistry in 76 lung cancer tissues from NSCLC patients. Expression of Twist and its correlation with HIF-1α, E-cadherin, and vimentin were analyzed. Small interfering RNA (siRNA) against Twist was used to knockdown Twist expression in hypoxic NSCLC cells, A549 and NCI-H460. Cellular invasion and protein levels of Twist, E-cadherin, and vimentin were evaluated by matrigel invasion assay and Western blot, respectively. Our results showed that in clinical samples, there was a significant association between Twist expression and differentiation degree, lymph node metastasis, and TNM stage. Correlation analysis demonstrated that expression of Twist was negatively correlated with E-cadherin expression, but positively associated with HIF-1α and vimentin expression. In cultured NSCLC cells, Twist messenger RNA (mRNA) and protein levels were upregulated under hypoxia, while knockdown of Twist suppressed potentiated invasion and expression of mesenchymal marker vimentin induced by hypoxia. Protein level of increased epithelial marker E-cadherin was shown along with Twist downregulation. These findings suggest that Twist promoting hypoxic invasion and metastasis of NSCLC may be associated with altered expression of EMT markers. Inhibition of Twist may be of therapeutic significance.

[Effects of lysyl oxidase down-regulation on invasion, migration and E-cadherin protein expression of hypoxic lung cancer cells].

OBJECTIVE: To observe the effects of lysyl oxidase (LOX) down-regulation on invasion, migration and epithelial-mesenchymal transition phenotype molecule E-cadherin protein expression, induced by hypoxia in lung cancer NCI-H460 cells. METHODS: Small interfering RNA against human LOX gene (LOX siRNA) was used to transfect lung cancer cells under normoxia (19%O2). After a 24 h incubation, the cells were plated for 24 h in hypoxic incubator (0.5%O2). Real-time polymerase chain reaction (PCR) was performed to detect the LOX mRNA expression. The protein levels of LOX and E-cadherin were determined by Western blot. And invasion and migration capacities were detected by transwell chamber. RESULTS: Compared with NCI-H460 cells under normoxia (set to 1), hypoxia increased to the levels of LOX mRNA and protein expression up to 26.04 ± 1.78 and 5.57 ± 1.27 respectively (both P < 0.05). Compared with control siRNA group (set to 1), LOX mRNA and protein expression after LOX siRNA transfection were 0.24 ± 0.03 and 0.29 ± 0.03 respectively, cellular invasive and migratory capacities were 0.57 ± 0.03 and 0.49 ± 0.02 respectively, the protein expression of E-cadherin was 2.17 ± 0.21 (all P < 0.05). CONCLUSION: LOX down-regulation reduces invasion and migration potentials of hypoxic human lung cancer cell and potentiates the protein expression of E-cadherin.

The modified shrinkage classification modes could help to guide breast conserving surgery after neoadjuvant therapy in breast cancer.

Purpose: The traditional shrinkage classification modes might not suitable for guiding breast conserving surgery (BCS) after neoadjuvant therapy (NAT). Aim was to explore the modified shrinkage classification modes to guide BCS after NAT. Methods: From April 2010 to 2018, 104 patients were included. All patients underwent MRI examinations before and after NAT. Residual tumors were removed and divided into more than 30 tissue blocks at 5-mm intervals. After performing routine procedures for paraffin-embedded histology, we made semiserial sections (6-µm thick). The MRI and pathology 3D models were reconstructed with 3D-DOCTOR software. Combined with traditional shrinkage modes and efficacy of NAT, we derived modified shrinkage classification modes which oriented by BCS purpose: modified concentric shrinkage modes (MCSM) and modified non concentric shrinkage modes (MNCSM). The MCSM means the longest diameter of residual tumor was less than 50% and ≤2cm in comparison with the primary tumor before NAT. Other shrinkage modes were classified as MNCSM. Results: According to traditional shrinkage modes, 50 (48.1%) cases were suitable for BCS;while 70 (67.3%) cases were suitable for BCS according to the modified shrinkage modes (p=0.007). The consistency of MRI 3D reconstruction in assessing modified shrinkage classification modes was 93.2%, while it was 61.5% when assessing traditional shrinkage modes. Multivariate analysis showed that primary tumor stage, mammographic malignant calcification, molecular subtypes and nodal down-staging after NAT were independent predictors of modified shrinkage modes (all p<0.05). A nomogram was created based on these four predictors. With a median follow-up time of 77 months, the recurrence/metastasis rate in the MCSM and MNCSM group was 7.1% and 29.4%, respectively. Conclusion: Modified shrinkage classification modes could help to guide the individualized selection of BCS candidates and scope of resection after NAT. MRI 3D reconstruction after NAT could accurately predict modified shrinkage modes and extent of residual tumor.

[Value of intraoperative GeneSearch(TM) BLN assay to detect breast cancer metastases in sentinel lymph nodes].

OBJECTIVE: To evaluate the value of GeneSearch(TM) BLN assay as an intraoperative diagnostic method of sentinel lymph node metastases in breast cancer patients. METHODS: Ninety consecutive patients were involved in this study. SLNs were intraoperatively identified and dissected, and then sectioned vertically to the long axis into multiple blocks. The odd blocks were tested by BLN assay and even ones prepared for frozen sectioning (FS), while all blocks were evaluated by touch imprint cytology (TIC). Post-operatively, residual tissues of the even blocks were assessed by histopathologic examination (4 - 6 µm thick serial sectioning permanent H&E slides were performed every 150 µm and one block made 6 slides). RESULTS: BLN assay could be performed within less than 35 min after learning curve of 10 cases. A correlation was found between cycle time values of mammaglobin or cytokeratin-19 and size of metastases, with Spearman correlation coefficients of 0.67 and 0.71, respectively. The accuracy, sensitivity, specificity, positive predict value (PPV) and negative predict value (NPV) of the assay were 95.6%, 93.3%, 96.7%, 93.3% and 96.7%, While FS had the sensitivity, specificity, PPV, NPV of 76.7%, 100%, 100%, 89.6%, and TIC of 73.3%, 100%, 100%, 88.2%, respectively. The sensitivity of the assay was higher than that of FS (P = 0.07), and was significantly higher than that of FS (P = 0.04). When assessing patients with micro-metastases, the assay had a sensitivity of 85.7%, which was significantly higher than that of FS and TIC (P = 0.03). CONCLUSION: GeneSearch(TM) BLN Assay can replace FS and TIC for the intraoperative assessment of SLN.

A Three lncRNA Set: AC009975.1, POTEH-AS1 and AL390243.1 as Nodal Efficacy Biomarker of Neoadjuvant Therapy for HER-2 Positive Breast Cancer.

PURPOSE: The study aimed to explore whether the expression of lncRNAs in primary tumors could predict nodal efficacy after neoadjuvant therapy (NAT) for HER2+ breast cancer. METHODS: Total RNA was extracted from HER2+ breast cancer tissues before NAT (n=103) and from 48 pairs of cancers and para-cancers tissues that did not receive NAT. Different lncRNAs were selected by microarray, validated by qPCR, and analyzed to illuminate their potential as nodal efficacy biomarkers after NAT. RESULTS: Our results demonstrated that three lncRNA sets, lncRNA-AL390243.1, POTEH-AS1, and lncRNA-AC009975.1, were up-regulated in non-apCR tissues. The AUC value was 0.789 (95%CI: 0.703-0.876). The multivariate logistic regression analysis identified the expression of lncRNA-AL390243.1 (OR 5.143; 95% CI: 1.570-16.847), tumor type (OR 0.144; 95% CI: 0.024-0.855), and nodal stage (OR 0.507; 95% CI: 0.289-0.888) as independent predictors for apCR after NAT in HER2+ patients (all p<0.05). Then the three predictors were used to create a predictive nomogram. The AUC value was 0.859 (95%CI: 0.790-0.929). The calibration curve showed a satisfactory fit between predictive and actual observation based on internal validation with a bootstrap resampling frequency of 1000. Patients with higher expression of lncRNA-AL390243.1 had worse survival. LncRNA-AL390243.1 was up-regulated more in the nodal positive subgroup than in the nodal negative subgroup (p=0.0271). CONCLUSION: The lncRNA-AL390243.1, POTEH-AS1, and lncRNA-AC009975.1 were upregulated in non-apCR breast cancer tissues. These three lncRNAs might have the potential to be used as predictive biomarkers of nodal efficacy of HER2+ breast cancer. Further studies are required to illuminate the underlying molecular mechanisms further.

Circulating Serum Exosomal Long Non-Coding RNAs FOXD2-AS1, NRIR, and XLOC_009459 as Diagnostic Biomarkers for Colorectal Cancer.

BACKGROUND: Exosomes derived from cancer cells encapsulate various kinds of tumor-specific molecules and thus can interact with adjacent or distant cells to mediate information exchange. Long non-coding RNAs (lncRNAs) in exosomes have the potential as diagnostic and prognostic biomarkers in different types of cancers. The current study was aimed to identify circulating exosomal lncRNAs for the diagnosis of colorectal cancer (CRC). METHODS: Exosomes were isolated from the serum by ultracentrifugation and verified by transmission electron microscope (TEM), qNano, and immunoblotting. Exosomal lncRNAs FOXD2-AS1, NRIR, and XLOC_009459 were selected by lncRNA microarray and validated by qPCR in 203 CRC patients and 201 healthy donors. The receiver operating characteristic curve (ROC) was used to assess the diagnostic efficiency of serum exosomal lncRNAs. RESULTS: Exosomal FOXD2-AS1, NRIR, and XLOC_009459 (TCONS_00020073) levels were significantly upregulated in 203 CRC patients and 80 early-stage CRC patients compared to 201 healthy donors, possessing the area under the curve (AUC) of 0.728, 0.660, and 0.682 for CRC, as well as 0.743, 0.660, and 0.689 for early-stage CRC, respectively. Notably, their combination demonstrated the markedly elevated AUC of 0.736 for CRC and 0.758 for early-stage CRC, indicating their potential as diagnostic biomarkers for CRC. CONCLUSIONS: Our data suggested that exosomal lncRNAs FOXD2-AS1, NRIR, and XLOC_009459 act as the promising biomarkers for the diagnostics of CRC and early-stage CRC.

The study of internal mammary lymph node dissection guided by modified radiotracer injection technique in breast cancer-a case report and review.

In addition to axillary lymph node (ALN), internal mammary lymph node (IMLN) could also provide important prognostic information. In this paper, we will introduce a case of breast cancer patient whose preoperative lymphoscintigraphy revealed that there were "hot-spots" in bilateral intercostal space. The bilateral IMLN found by preoperative lymphoscintigraphy is a rare phenomenon. She received ipsilateral internal mammary sentinel lymph node biopsy (IM-SLNB) and IMLN dissection and contralateral IM-SLNB. She was diagnosed as pT2N3bM1 breast cancer based on the positive IMLN and positive ALN. After performing surgery, the pathology indicated: (left breast) invasive ductal carcinoma (3.0×3.0 cm2), ALN (3/30), ipsilateral internal mammary sentinel lymph node (IMSLN) (1/2), IMLN (0/2); contralateral IMSLN (1/1). After performing IMLN surgery, the pathology staging increased from pT2N1aM0 to pT2N3bM1. And the irradiation therapy choice had been changed, she received irradiation therapy include chest wall, supraclavicular region, ipsilateral IMLN and contralateral IMLN. The treatment benefit had been increased. When the ipsilateral internal mammary lymphatic vessels were obstructed, deep lymphatic system might drain from ipsilateral IMLN to contralateral IMLN. The contralateral IMLN metastasis belongs to distant metastasis. The IMLN irradiation therapy should be tailored and balanced based on the statues of IMLN. With effective application of systemic therapy, the localized treatment advantage benefited from IMLN surgery might be transferred to survival benefit.

Circulating serum exosomal miR-20b-5p and miR-3187-5p as efficient diagnostic biomarkers for early-stage non-small cell lung cancer.

IMPACT STATEMENT: The high mortality of non-small cell lung cancer (NSCLC) is mainly because the cancer has progressed to a more advanced stage before diagnosis. If NSCLC can be diagnosed at early stages, especially stage 0 or I, the overall survival rate will be largely improved by definitive treatment such as lobectomy. We herein validated two novel circulating serum ExmiRs as diagnostic biomarkers for early-stage NSCLC to fulfill the unmet medical need. Considering the number of specimens in this study, circulating serum exosomal miR-20b-5p and miR-3187-5p are putative NSCLC biomarkers, which need to be further investigated in a larger randomized controlled clinical trial.

[Down-regulation of TopI by RNA interference reduces the drug-sensitivity of small cell lung cancer cell lines to topotecan].

OBJECTIVE: To investigate the expressions of TopI gene in small cell lung cancer cell line H446, and explore the influence of TopI on the chemosensitivity of the cell line to topotecan (TPT). METHODS: Western blot was performed to detect the TopI expression in H446 cells. Lipofectamine 2000 was used for the transient transfection of H446 cells by siRNA, and the transfection efficacy was detected. TopI mRNA was analyzed by quantitative RT-PCR and TopI protein was detected by Western blot to selected effective siRNA. The drug-sensitivity to topotecan (TPT) was evaluated by MTT assay. RESULTS: TopI gene was expressed in H446 cells. Lipofectamine 2000 mediated the siRNA effectively (88.67%). Compared with its parental cells, RT-PCR results showed that TopI mRNAs in transfected cells were reduced by (95.7 +/- 1.6)%, (90.8 +/- 1.6)%, (96.1 +/- 2.7)% and (96.3 +/- 1.8)%, respectively, and decreased significantly at protein level. By MTT assay, the inhibition rate of TPT to H446 cells transfected by siRNA was lower than that of control group at same concentrations (P < 0.01). CONCLUSION: siRNAs can silence the expression of TopI and decrease the drug-sensitivity of H446 cells to TPT.

[Molecular imaging of epidermal growth factor receptor in glioma-bearing rats].

OBJECTIVE: To investigate the value of 11C-PD153035 as an EGFR imaging agent in C6 tumor-bearing rat. METHODS: The tumor-bearing rats were generated by subcutaneous injection of glioma C6 cells. Positron emission tomography/computer tomography (PET/CT) scans started as soon as intravenous injection of 11C-PD153035 (15-20 MBq/0.3 ml) was completed, images were collected continuously. The region of interest (ROI) was used to study the percentage of radioactivity in major organs and implanted tumors in the rats. The accumulation and blocking study in vitro was completed. RESULTS: There were significant differences in 11C-PD153035 uptake among major organs. The maximum uptake in the organs ranked in the following order: liver > gastrointestinal tract > kidney > lung > brain > muscle. Radioactivity could be also observed in the tumors. The radioactivity ratio (T/NT, target/non-target) peaked (4.15) at 40 - 50 min post injection. The in vitro blocking study showed that 11C-PD153035 uptaken by C6 cells could be blocked by PD153035. CONCLUSION: The results of this study show that 11C-PD153035 can be uptaken by EGFR-expressing tumors. 11C-PD153035 has a potential as a bioprobe to yield useful information for both diagnosis and therapy of tumors. However, the high concentration of 11C-PD153035 in the gastrointestinal tract is unfavorably affecting the tumor detection in these organs.

Further characterization of the epidermal growth factor receptor ligand 11C-PD153035.

BACKGROUND: (11)C-4-N-(3-bromoanilino)-6,7-dimethoxyquinazoline ((11)C-PD153035) has been reported as a tracer for imaging human tumors that overexpress epidermal growth factor receptor (EGFR). However it is still unclear whether (11)C-PD153035 uptake correlates with EGFR expression levels. The objective of this study was to investigate the relationship between (11)C-PD153035 accumulation and EGFR expression levels. METHODS: Synthesis of (11)C-PD153035 was performed in the Tracerlab FXc system. Accumulation of (11)C-PD153035 by MDA-MB-468, A549 and MDA-MB-231 cells was measured in vitro. There were six tumor-bearing mice in each group. (11)C-PD153035 uptake in tumors was determined by positron emission tomography/computed tomography (PET/CT). Tumor/normal muscle tissue (T/NT) analysis in PET images was applied to quantify the PET data. Sixty minutes after PET/CT scanning, the nude mice were sacrificed and the tumors were excised. The (11)C-PD153035 accumulation in different tumors was determined by a gamma counter. RESULTS: Close correlation existed between the uptake and the level of EGFR expression both in vitro and ex vivo (r(2) = 0.72, P < 0.001; r(2) = 0.63, P = 0.003). When the static T/NT analysis method was applied to analyze the PET data, the observed correlation was again excellent (r(2) = 0.70, P = 0.001). CONCLUSIONS: The uptake of PET tracer (11)C-PD153035 closely correlates with the EGFR expression levels in tumor cells. (11)C-PD153035 has the potential to yield useful information for both cancer diagnosis and therapy.

[Prolonged hypoxia induces GST-pi expression and drug resistance in lung adenocarcinoma cell line SPCA1].

OBJECTIVE: To investigate the association of hypoxia with GST-pi expression in lung adenocarcinoma cells SPCA1 and the drug resistance in hypoxic condition. METHODS: RNA and protein in SPCA1 cells treated with hypoxia (0.5% O(2)) for 16 h were isolated. HIF-1alpha and GST-pi mRNA expression was detected by real time RT-PCR, and HIF-1alpha protein was detected by Western blotting analysis. GST-pi protein was finally analyzed on a FACScan flow cytometer using Cellquest software. Sensitivity of Doxorubicin and Mitomycin in hypoxic SPCA1 cells was detected using cell clonogenic assay. RESULTS: HIF-1alpha expression was up-regulated on mRNA and protein levels in hypoxic SPCA1 cells, so as was GST-pi expression. Doxorubicin resistance was increased, but hypoxia had no significant influence on Mitomycin resistance. GST-pi expression had no significant change after HIF-1alpha was down-regulated by RNA interfering. CONCLUSION: Though prolonged hypoxia can induce GST-pi expression and increase drug resistance such as to Doxorubicin, HIF-1alpha expression has no correlation with GST-pi expression.

Regulation of the chemokine receptor CXCR4 and metastasis by hypoxia-inducible factor in non small cell lung cancer cell lines.

Hypoxia promotes metastatic potential of tumor cells by largely unknown mechanisms. Hypoxia inducible factor (HIF) is a heterodimeric transcription factor consisting of alpha and beta (ARNT) subunits and plays an important role in tumor microenvironment. CXCR4 is a cell surface receptor that has been shown to mediate the metastasis of various tumors. CXCR4 induction by hypoxia is dependent on both activation of HIF and transcript stabilization. To investigate the mechanisms involved in hypoxia-induced metastasis and hypoxia-mediated chemokine receptor CXCR4 expression, we used lentiviral vector mediated RNA interfering (RNAi) to knock down expression of HIF-1alpha or HIF-2alpha in two NSCLC cell lines to investigate HIF-dependent invasion, migration and adhesion. Here we show that: (1) hypoxia is an important factor in regulating CXCR4 mediated metastasis and the cells exhibited reducing invasion, adhesion and migration in response to CXCL12 after knocking down HIF. (2) HIF-1alpha and HIF-2alpha are essential for hypoxic cellular response to cancer invasion and adhesion through upregulation of CXCR4. HIF-1alpha and HIF-2alpha are playing important roles in tumor metastasis, which may offer for future intervention strategies. We also show that the lentivirus mediated RNAi technology is very effective on knocking down gene expression.

[Expression of MDR1 and GST-pi in osteosarcoma and soft tissue sarcoma and their correlation with chemotherapy resistance].

OBJECTIVE: To explore the expression of multidrug resistance gene 1 ( MDR1), glutathione-S-transferases-pi (GST-pi) in osteosarcoma and soft tissue sarcoma tissues from 34 patients and their correlation with chemotherapy resistance. METHODS: MDR1 and GST-pi expressions were analyzed by real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) and flow cytometry (FCM) at mRNA and protein levels, respectively. Chemotherapy sensitivity on adriamycin, cisplatinum, fluorouracil, mitomycin C, dacarbazine, vincristine, methotrexate in tumor tissues were detected by MTT assay. RESULTS: The nonsensitive rates on adriamycin, cisplatinum, fluorouracil, mitomycin C, dacarbazine, vincristine, methotrexate in tumor tissues were 41.18%, 17.7%, 47.1%, 50.0%, 76.5%, 61.8% and 52.9%, respectively. The expression of P-glycoprotein (P-gp) and GST-pi in tumor tissues was 1.54 and 2.58 (relative fluorescence intensity). Chi2 analysis showed that there was a positive correlation between P-gp expression and drug resistance on ADM, GST-pi expression and resistance on ADM, DDP and MMC (P < 0.05). There was not seen obvious correlation between expression of MDR1, GST-pi and age, gender, pathological type, tumor size in osteosarcoma and soft tissue sarcoma patients (P > 0.05). The expression of GST-pi was increased in patients receiving preoperative chemotherapy. The rate of postoperative recurrence was higher in patients with higher GST-pi expression level than those with lower GST-pi expression level before operation (P < 0.05). CONCLUSION: Individual differences exist in chemotherapy sensitivity and expression of MDR1 and GST-pi in osteosarcoma and soft tissue sarcomas patients. Chemotherapy can induce up-regulation of GST-pi protein expression. Primary high expression of GST-pi is the main mechanism of resistance of osteosarcoma and soft tissue sarcomas to chemotherapy and is related to poor prognosis.

[The clinical significance of P-glycoprotein, lung resistance protein and multidrug resistance related protein expressions in infiltrating breast carcinoma tissues].

OBJECTIVE: Our purpose was to explore clinical significance of three kinds of drug resistance related proteins, including P-glycoprotein (P-gp), lung resistance protein (LRP), multidrug resistance related protein (MRP) expressions in infiltrating breast carcinoma tissues. METHODS: Flow cytometry was used to analyze the expression of drug resistance related proteins in carcinoma tissues and corresponding para-carcinoma tissues from 68 primary infiltrating breast carcinoma patients. RESULTS: The relative fluorescence intensity (RFI) expressed by median (M) of P-gp, LRP and MRP in breast carcinoma tissues were 0.58, 0.51and 0.56 respectively. In corresponding para-carcinoma tissues, their expression level were 0.26, 0.33 and 0.30. Their difference were obvious (P < 0.05). There were not notable differences of drug resistance related protein expressions in different ages, different tumor sizes, different pathological types, different clinical stages, estrogen receptor or progestin receptor negative and positive patients (P > 0.05). The expressions of P-gp and MRP in lymph node metastasis positive patients were 0.61and 0.69, in lymph node metastasis negative patients they were 0.54 and 0.45. There were no notable differences of P-gp and MRP expression in lymph node metastasis negative and positive patients. The expression of LRP (M = 1.49) was higher in lymph node metastasis positive patients than that in negative patients (M = 0.50, P < 0.05). The correlation analysis showed that there was positive correlation between the expression of LRP and MRP in breast carcinoma tissues (P < 0.05). CONCLUSION: The clinical significance of three kinds of drug resistance related protein expressions had differences in infiltrating breast carcinoma tissues. The expression of LRP had related with lymph node metastasis status and had positive correlation with MRP expression. It can become a target in forecasting breast carcinoma prognosis.

Preparation study of indocyanine green-rituximab: A new receptor-targeted tracer for sentinel lymph node in breast cancer.

An appropriate receptor-targeted tracer for sentinel lymph node biopsy (SLNB) was prepared. We combined the fluorescence tracer (Indocyanine green, ICG) with Rituximab (a chimeric human/murine monoclonal antibody targeting the CD20 antigen on the surface of lymphocyte) directly to produce a new tracer (ICG-Rituximab). When the new tracer drains to the lymph node, Rituximab will combine with CD20 receptor on the B-cell surface in the lymph node. If the statue of antibody-receptor connection does not reach saturation, the number of Rituximab is less than CD20. With this appropriate injection dose, the new tracer could only stay in sentinel lymph node (SLN) and make it imaging. Positive fluorescence SLN was detected 12 minutes after injection with no other organs imaging. The imaging of SLN was stable and clear for 20-24 hours. Due to SLN stained with more ICG than the lymphatic vessel, the fluorescence situation of SLN would be brighter than the vessel. The surgeon can detect the positive fluorescence SLN easily without following the fluorescence imaging lymphatic vessel. The results of our preliminary study showed that the new tracer might be useful for improving SLN imaging and worth further clinical study. SLNB with the new tracer could be a convenient method for detecting SLN and would become a standard performance in clinical practice.