e16514 Background: Antiangiogenic therapy combined with immunotherapy significantly improved the progression-free survival (PFS) and preliminary survival benefit for advanced clear-cell renal cell carcinoma (ccRCC). However, no combined immune therapies are currently approved for ccRCC in China. Fruquintinib (F, a highly selective VEGFR inhibitor) combined with sintilimab (S, an anti-PD-1 monoclonal antibody) showed encouraging antitumor activity in multiple solid tumors. Here, we reported the results of F plus S in metastatic ccRCC cohort from an open-label, single-arm phase 2 study (NCT03903705). Methods: Patients (pts) with pathologically confirmed metastatic ccRCC who were treatment-naive or had failed previously first-line treatment were eligible. They received F (5 mg, 2 weeks on/1 week off, orally, QD) plus S (200 mg, IV, Q3W) in 21-day cycles until disease progression or unacceptable toxicity; treatment of S was allowed for up to 24 months. The primary endpoint was objective response rate (ORR) per RECIST v1.1. Results: As of cutoff (November 30, 2022), a total of 42 pts (22 treatment-naïve pts, and 20 previously first-line treated pts) were enrolled and received the combination treatment. Median (range) age was 59.0 (34.2-72.4) years; 34 (81.0%) pts were male. Among previously treated pts, 19 (19/20, 95.0%) pts had received VEGFR therapy as prior first-line treatment, including sunitinib (3/20, 15.0%), pazopanib (7/20, 35.0%), axitinib (2/20, 10.0%) and others (7/20, 35.0%). International Metastatic RCC Database Consortium (IMDC) risk included: favorable in 15 (35.7%) pts, intermediate in 22 (52.4%) pts, and poor in 5 (11.9%) pts. The median follow-up duration was 14.8 months (m) and 23.3 m for treatment-naive and previously treated pts, respectively. All pts had at least 1 post-baseline tumor assessment. Confirmed ORR was 68.2% and 60.0%, median duration of response was not reached (NR) and 13.9 m, and disease control rate was 95.5% and 85.0% for treatment-naive and previously treated pts, respectively. Median PFS (mPFS) was NR and 12m-PFS rate was 64.8% in treatment-naive pts; mPFS was 15.9 m in previously treated pts. The median treatment duration of F and S was both 14.7 m. All pts experienced at least 1 treatment-emergent adverse event (TEAE). Common (≥5%) grade ≥3 TEAEs were hypertension (21.4%), hypertriglyceridaemia (9.5%), amylase increased (7.1%), cerebral infarction (7.1%) and proteinuria (7.1%). No new safety signals were observed. Conclusions: F plus S demonstrated a favorable efficacy with manageable toxicity for treatment-naive and previously first-line treated metastatic ccRCC. A phase 2/3 study is being conducted to confirm this combination efficacy in the second-line treatment of ccRCC (NCT05522231). Clinical trial information: NCT03903705 .
Objective To study the impact of TDGF-1 gene silience by small interfering RNA(siRNA)on the invasion and migration of human breast cancer cell. Methods 3 siRNA fragments were designed according to the characteristic of TDGF-1 gene sequence and the most appropriate siRNA was selected by fluorescence real-time quantitative RT-PCR method. After the human breast cancer cell line MDA-MB-468 was transfected by the selected TDGF-1 siRNA, mRNA and protein of TDGF-1 were determined by real time quantitative RT-PCR and western blot respectively. The migration and invasion ability of the cancer cell were evaluated by wound-healing assay and Boyden chamber model respectively. Results siRNA could down-regulate the level of mRNA and protein of TDGF-1 in a dose-and time-dependent manner. In vitro experiment showed that TDGF-1 siRNA transfection can effectively inhibit the clonal growth, invasion and migration of breast cancer cell in a dose-dependent manner. Conclusions TDGF-1 gene may play an important role in the migration and invasion of human breast cancer cells. siRNA transfection can inhibit the invasion of human breast cancer cells.
Key words:
Breast carcinoma; Teratocarcinoma-derived growth factor-1; Invasion; Migration; Small interfering RNA
Objective To study the effects of teratocarcinoma-derived growth factor-1 (TDGF-1) gene small interfering RNA (siRNA) on adhesion and invasion of human melanoma cell.Methods Realtime fluorescent quantitative polymerase chain reaction(FQ-PCR) was used to evaluate the TDGF-1 mRNA expression of human melanoma cell lines A-375,C-918 and M14.The highest expression of TDGF-1 was transfected with different dose of TDGF-1 siRNA.The expression of TDGF-1 mRNA and protein were were determined by Real-time quantitative PCR and Western blotting,respectively.Cell adhesion was exmined by methyl thiazol tetrazolium (MTT) assay,and cell invasion was evaluated by boyden chamber,respectiv ely.The matrix metalloproteinase-13 (MMP-13) were examined by enzyme linked immunosorbent assay (ELISA) assay.Results The results from FQ-PCR showed that the TDGF-1 mRNA of A-375,C-918 and M14 cell line is 0.589 ±0.081,0.712 ±0.065,and 1.517 ±0.085.After M14 cell line was transfected by TDGF-1 siRNA,the results of the MTT assay showed that the A Values of Con-A,Con-B,3.125 nmol/L,6.250 nmol/L,and 12.500 nmol/L siRNA were 0.89 ±0.15,0.85 ±0.12,0.62 ±0.09,0.51 ±0.08,0.33 ± 0.06,respectively (P ＜ 0.05) ; the results of Boyden assay showed that the membrane cell number of Con-A,Con-B,3.125 nmol/L siRNA,6.250 nmol/L siRNA,and 12.500 nmol/L siRNA were 37.8 ± 1.6,36.9±1.5,21.8±1.2,11.6±0.8,and 5.6±0.5,respectively (P＜0.05).The results from ELISA showed that urokinase-type plasminogen activator (uPA) content of Con-A,Con-B,3.125 nmol/L siRNA,6.250 nmol/L siRNA,and 12.500 nmol/L siRNA were (85.2 ± 1.8),(84.8 ± 1.5),(51.6 ± 1.2),(35.6 ± 0.8),(17.8 ± 0.6) ng/L.Conclusion TDGF-1 gene might play an important role in adhesion and invasion of human melanoma cancer cell.The siRNA targeted TDGF-1 could effectively inhibit adhesion and invasion of human melanoma cancer cell through downregulation uPA.
Key words:
Melanoma ; Teratocarcinoma-derived growth factor-1 ; Adhesion ; Invasion ; Urokinase-type plasminogen activator
Additional file 2: Figure S1. The effect of CDDP on cell viability of ESCC cells. a The effect of CDDP on cell viability of KYSE30/CDDP-R and KYSE30 cells. b The effect of CDDP on cell viability of TE1/CDDP-R and TE1 cells. Figure S2. Representative images of RNA FISH of NORAD in TE1/CDDP-R and TE1 cells (× 1000), which show that NORAD is predominantly located in the cytoplasm. Nuclei are stained with DAPI. Figure S3. The effect of sh-NORAD and overexpression vector on the expression of NORAD and proliferation ability of ESCC cells. a The effect of sh-NORAD on NORAD expression in KYSE30/CDDP-R and TE1/CDDP-R cells. Error bars denote SD of triplicates. ***P
Abstract Analogous to DNA methylation and histone modification, RNA modification, as another epigenetic layer, plays an important role in many diseases, especially in tumours. As the most common form of RNA modification, m 6 A methylation has attracted increasing research interest in recent years. m 6 A is catalysed by RNA methyltransferases METTL3, METTL14 and WTAP (writers), m 6 A is removed by the demethylases FTO and ALKBH5 (erasers) and interacts with m6A-binding proteins, such as YT521-B homology (YTH) domain-containing proteins. This article reviews recent studies on methylation modification of m 6 A in gastrointestinal tract cancers.
To compare the differences in uptake of 2-deoxy-D-glucose (2-DG)-conjugated nanoparticles between breast carcinoma MDA-MB-231 cells with high metabolism and breast fibroblasts with normal metabolism, and investigate the feasibility of using the coated nanoparticles as a MRI-targeted contrast agent for highly metabolic carcinoma cells.The γ-Fe2O3@DMSA-DG was prepared. The glucose metabolism level of both cell lines was determined. The targeting efficacy of γ-Fe2O3@DMSA-DG and γ-Fe2O3@DMSA NPs to breast carcinoma MDA-MB-231 cells and breast fibroblasts at 10 min, 30 min, 1 h and 2 h was measured with Prussian blue staining and UV colorimetric assay. MRI was performed to visualize the changes of T2WI signal intensity.Prussian blue staining showed more intracellular blue granules in the MDA-MB-231 cells of γ-Fe2O3@DMSA-DG NPs group than that in the γ-Fe2O3@DMSA NPs group, and the γ-Fe2O3@DMSA-DG uptake was greatly competed by free D-glucose. As revealed by UV colorimetric assay, MDA-MB-231 cells also showed that the cellular iron amount of γ-Fe2O3@DMSA-DG group was significantly higher than that of the γ-Fe2O3@DMSA group and γ-Fe2O3@DMSA-DG + D-glucose group, statistically with a significant difference between them. MRI showed that the signal intensity of γ-Fe2O3@DMSA-DG group was decrease significantly, the T2 signal intensity was decreased by 10.5%, 37.5%, 72.9%, 92.0% for 10 min, 30 min, 1 h and 2 h, respectively. In contrast, the signal intensity did not show obvious decrease in the γ-Fe2O3@DMSA-DG group, the T2 signal intensity was decreased by 8.5%, 11.4%, 32.0%, 76.7% for 10 min, 30 min, 1 h and 2 h, respectively. However, HUM-CELL-0056 cells did not produce apparent difference for positive staining in the γ-Fe2O3@DMSA-DG group, γ-Fe2O3@DMSA group and γ-Fe2O3@DMSA-DG+D-glucose group, and the signal intensity also did not produce apparent difference.γ-Fe2O3@DMSA-DG has good targeting ability to highly metabolic breast carcinoma (MDA-MB-231) cells. It is feasible to serve as a specific MRI-targeted contrast agent for highly metabolic carcinoma cells, and deserves further studies in vivo.
Background: The positive results of the apatinib phase III trial have cast new light on treatment for patients with advanced gastric cancer (GC). However, in terms of safety, apatinib toxicities may lead to a dose modification or treatment interruption. Therefore, properly intervene is urgently needed to reduce toxicities and help patients benefit from apatinib treatment.Methods: The in vitro and in vivo assays were conducted to investigate the mechanisms of apatinib in GC. Autophagy functional assays were performed. MicroRNA sequencing and circular RNA sequencing of the GC tumor xenografts from the control and apatinib groups were performed to further study the mechanism.Results: In this study, we found that apatinib inhibited the growth of GC cells and xenograft tumors. Apatinib promoted autophagy activation via upregulation of ATG7 expression, and autophagy inhibition enhanced apatinib-induced apoptosis. With microRNA and circular RNA sequencing analyses of GC xenograft models, we demonstrated that circRACGAP1 functioned as an endogenous sponge for miR-3657 to inhibit its activity and further upregulate ATG7 expression. Silencing of circRACGAP1 inhibited apatinib-induced autophagy, which was rescued by miR-3657. Moreover, knockdown of circRACGAP1 sensitized GC cells to apatinib via autophagy inhibition in vitro and in vivo.Conclusions: These findings provided the first evidence that the circRACGAP1-miR-3657-ATG7 axis mediates a novel regulatory pathway critical for the regulation of autophagy and apatinib sensitivity in GC cells and xenografts. Thus, specific blockage of circRACGAP1 may be a potential therapeutic strategy to reduce the toxicities of apatinib and enhance its therapeutic effect in human GC.Funding Statement: This study was supported by grants from the National Natural Science Foundation of China (No. 81672896 and No. 81520108027), the National Key Research and Development Program: the key technology of palliative care and nursing for cancer patients (ZDZX2017ZL-01), the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) (JX10231802), and Jiangsu Provincial Key Research and Development Special Fund (BE2015666).Declaration of Interests: The authors declare no conflict of interest.Ethics Approval Statement: All experiments were carried out according to the National Institutes of Health Guidelines for the Care and Use of Laboratory Animals (NIH Publication No. 80-23) revised in 1996. The animal studies were reviewed and approved by the Ethics Committee for Animal Studies at Nanjing Medical University.
Background Preoperative albumin-globulin ratio (AGR) reflects both malnutrition and systemic inflammation in cancer patients. In particular, systemic inflammation has been reported to contribute to tumor progression and poor oncological outcome in various malignancies. However, the prognostic value of preoperative AGR in upper tract urothelial carcinoma (UTUC) has not been examined. Methods We retrospectively reviewed medical data of 187 operable UTUC patients in a Chinese cohort with a high incidence of chronic kidney disease (CKD). AGR was calculated as [AGR = albumin/(serum total protein—albumin)]. The associations of preoperative AGR with clinicopathologic characteristics and prognosis were assessed. Multivariate analyses using Cox regression models were performed to determine the independent prognostic factors. Results The median (IQR) preoperative AGR was 1.50 (1.30–1.70), and the optimal cutoff value was determined to be 1.45 according to the receiver operating curve analysis. Low AGR was significantly associated with female gender, high CKD stage and tumor grade (P < 0.05). Eighty-three patients died before the follow-up endpoint. Kaplan-Meier analysis showed that an AGR < 1.45 predicted significantly poorer overall and cancer-specific survivals compared to an AGR ≥ 1.45 (P < 0.001 and P = 0.008, respectively). Multivariate analyses showed that an AGR < 1.45 was an independent risk factor for poorer overall and cancer-specific survivals (P = 0.002 and P = 0.015, respectively). Conclusions Preoperative AGR can act as an effective biomarker with easy accessibility for evaluating the prognosis of patients with UTUC. AGR should be applied in UTUC patients for risk stratification and determination of optimal therapeutic regimens.
Prealbumin is a sensitive indicator of liver function and nutritional status. This meta-analysis aimed to examine the association of the serum prealbumin level with the prognosis of patients with hepatocellular carcinoma (HCC) undergoing hepatectomy. We comprehensively searched the PubMed, Embase, Wanfang, China Academic Journals (CNKI), and SinoMed databases up to September 1, 2021. Eligible studies should report the association of the serum prealbumin level with prognosis and provide the multivariable-adjusted risk estimates of the outcomes of interest in HCC patients undergoing hepatectomy. A total of 11 studies with 7,442 HCC patients were identified and analyzed. Meta-analysis of a fixed effects model showed that a low serum prealbumin level was associated with poor overall survival [hazard ratio (HR) = 1.54, 95% confidence interval (CI) = 1.42-1.68], recurrence-free survival (HR = 1.34, 95% CI = 1.17-1.52), and a higher risk of postoperative hepatic insufficiency (HR = 2.21; 95% CI = 1.36-3.60) in HCC patients. Sensitivity and subgroup analyses confirmed the robustness of low serum prealbumin in predicting poor overall survival. This meta-analysis indicated that a low preoperative serum prealbumin level was significantly associated with adverse prognosis in HCC patients undergoing hepatectomy.
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