Chemo-photothermal immunotherapy for eradication of orthotopic tumors and inhibition of metastasis by intratumoral injection of polydopamine versatile hydrogels

Cancer remains one of the leading causes of death globally and metastasis always leads to treatment failure.Here,we develop a versatile hydrogel loading photothermal agents,chemotherapeutics,and immune-adjuvants to eradicate orthotopic tumors and inhibit metastasis by combinational therapy.Hydrogel networks were synthesized via the thiol-Michael addition of polydopamine(PDA)with thiolated hyaluronic acid.PDA acted as a cross-linking agent and endowed the hydrogel with excellent photothermal property.Meanwhile,a chemotherapeutic agent,doxorubicin(DOX),was loaded in the hydrogel viaπ-πstacking with PDA and an immune-adjuvant,CpG-ODN,was loaded via electrostatic interaction.The release of DOX from the hydrogel was initially slow but accelerated due to near infrared light irradiation.The hydrogels showed remarkably synergistic effect against 4T1 cancer cells and stimulated plenty of cytokines secreting from RAW264.7 cells.Moreover,the hydrogels eradicated orthotopic murine breast cancer xenografts and strongly inhibited metastasis after intratumoral injection and light irradiation.The high anticancer efficiency of this chemo-photothermal immunotherapy resulted from the strong synergistic effect of the versatile hydrogels,including the evoked host immune response.The combinational strategy of chemo-photothermal immunotherapy is promising for highly effective treatment of breast cancer.

Biodistribution and Anti-tumor Activities of the ^(131)I-labeled Rituximab in Nude Mice Bearing Human Burkitt's lymphoma

OBJECTIVE To explore the biodistribution and anti-tumoractivity of ^(131)I labeled rituximab injected intratumorally orintraperitoneally in vivo in nude mice bearing Raji human Burkitt's lymphoma xenografts.METHODS The rituximab and the mouse IgG were labeled withNa^(131)I using the IODO-GEN method.BALB/C nude mice werexenografted with ^(131)I-Rituximab or ^(131)I-IgG and killed on the 1st,3rd,7th,and 15th day after injection.The tumor/non-tumor ratio(T/NT)and the dose injected in each gram of the tissue(%ID/g)from12 organs or tissues of interest,e.g.tumor,blood,were calculated.The long and short axes of each tumor were measured by calipersat 2-3-day intervals after treatment,and the growth inhibition ofthe tumor was calculated using the MIRD formula.RESULTS When comparing intraperitoneal injection(IP)andintratumoral injection(IT)of ^(131)I-IgG,intratumoral injection of^(131)I-rituximab produced a significantly higher tumor/non-tumorratio in all tissues and organs of interest on the 1st,3rd,and 7thday,respectively(P<0.05).The %ID/g of tumor was 1.4-1.7-foldand 1.5-3.7-fold in the IP and IgG IT groups,respectively,but the%ID/g of non-tumors was significantly lower in the IP group andIgG IT group.Similarly,the tumor growth was greatly inhibitedby intratumoral injection of the ^(131)I-rituximab,whereas it wasless inhibited by other forms of the treatment(P<0.05).However^(131)I-rituximab injected intratumorally inhibited tumor growth ina dose-dependent manner.The inhibition rate was less with alow dose(75μCi)and greater with a high dose(150μCi),yet thedifference was not significant(P>0.05).CONCLUSION Tumors can absorb the highest amount of theradiolabelled antibodies,and the tumor/non-tumor ratios in thegroup with intratumoral injection of the ^(131)I-rituximab resulted inthe optimal anti-tumor activity.

Inhibition of lung cancer stem cells self-renewal and tumorigenicity by lentivirus-delivered Bmi1 shRNA

Objective： The aim of the study was to observe the effect of Bmil reduction on the self-renewal and tumorigenicity ability of lung cancer stem cells （LCSCs） in human lung adenocarcinoma. Methods： Human lung adenocarcinoma cells A549 were consecutively passaged in NOD/SCID mice treated with Paclitaxel weekly. The proportions of LCSCs in A549 cells and the cells from the third passage （A549-3rd） were compared. The expression of Bmil in LCSCs was silenced by intratumoral injection with lentivirus-delivered Broil small hairpin RNA （shRNA）. RT-PCR and Western blot were used to test the mRNA and protein expressions of Broil in LCSCs. The protein level of p16INK4A was analyzed by Western blotting. The self- renewal and tumorigenicity ability of LCSCs were evaluated by counting the sphere formation rate in serum-free medium and the tumor formation rate in NOD/SCID mice. Results： In vivo passaging ofA549 cells under chemotherapy pressure enriched for LCSCs. The expression of Broil in LCSCs increased. Down-regulation of Bmil by RNA interference resulted in reduced self-renewal and tumorigenicity ability of LCSCs and paralleled the increased expression of p16INK4A, a Bmil target. Conclu- sion： Broil regulates self-renewal and tumorigenicity of LCSCs by silencing some target genes, including p16INK4A.

Experimental Research of Tankyrase1 Antisense Oligodeoxynucleotides on the Proliferation of Lung Cancer Cell Nodules

OBJECTIVE To observe the effects of sense and antisense oligodeoxynucleotides of tankyrase 1 （TANK1-SODN and TANK1-ASODN） on murine tumor growth following intratumoral injection, investigate the actual suppressing result and mechanism of TANK1-ASODN on cancer cell proliferation, and discuss the possibility of using it in gene therapy on human lung cancer cells. METHODS After BALB/c nude mice had been subcutaneously inoculated with human lung cancer cell line CALU and it had grown into tumor nodules, we distributed these mice randomly into 3 groups： 4 in saline treatment group, and 5 each in TANK1-SODN group, and TANK1-ASODN groups. Then multiple direct intratumoral injections of synthesized TANK1-ASODN given continuously into tumor nodules for 16 days, this was compared with TANK1-SODN and saline control groups. During the experiment we measured the tumor volume every 5 days with vernier calipers; observed the histopathological characteristics of tumor tissues under microscope; went further to detect the minute changing of ultrastructure of cancer cells by electron microscope; tested the expression levels of ki67 and hTERT protein by means of SABC immunohistochemical method; and detected the lung cancer cells＇ hTERT mRNA expression level by hybridization in situ （ISH） in each group. RESULTS After 16 days of continuous injection, the tumor volume in TANK1-ASODN group was significantly smaller than the other 2 groups （both P 〈 0.01）; quite a lot of tumor cell degeneration and necrosis were observed in mice given TANK1-ASODN. The results of electron microscope also showed that TANK1-ASODN has the power to kill cancer cells in various ways. Moreover, statistically signi.cant decreases in the positive expression ratio of Ki67 Labeling Index （P 〈 0.01）, hTERT protein （P 〈 0.01）, and hTERT mRNA （P 〈 0.01） were consistently observed in the TANK1-ASODN group. CONCLUSION Human lung cancer cell line CALU expressed high telomerase activity. TANK1-ASODN had the ability to decline the high expression level of hTERT; inhibit the activity of telomerase, accelerate tumor cell degeneration and necrosis; and then suppress the proliferation of cancer cells.

Efficacy of Aidi Injection(艾迪注射液)on Overexpression of P-Glycoprotein Induced by Vinorelbine and Cisplatin Regimen in Patients with Non-Small Cell Lung Cancer

Objective：To investigate the efficacy of Aidi Injection（艾迪注射液） on overexpression of P-glycoprotein（P-gp） induced by vinorelbine and cisplatin（NP） regimen in patients with non-small cell lung cancer（NSCLC）, and study the difference between intravenous administration and targeting intratumor administration of Aidi Injection with thoracoscope. Methods：Totally 150 patients with NSCLC were randomly assigned to the control group, the intravenous group and the intratumor group by the random envelope method, 50 cases in each group. The patients were treated with NP regimen（2 cycles）, NP regimen（2 cycles） plus Aidi intravenous injection, or NP regimen（2 cycles） plus Aidi intratumor injection with thoracoscope, respectively for 6 weeks. The clinical efficacy was observed based on Response Evaluation Criteria in Solid Tumors（RECIST） rules, the expression of P-gp in the tumor tissue was tested before, 3 and 6 weeks after treatment, the safety was evaluated by monitoring the toxicity in the process of treatment, and the progressionfree survival（PFS） was measured. Results：Fifteen cases dropped out because of the irreconcilable conditions which had no relationship with the treatment, 4 in the control group, 5 in the intravenous group, and 6 in the intratumor group, respectively. Compared with the control group, the response rates（complete remission ＋ partial response） and the disease control rates（complete remission ＋ partial response ＋ stable disease） were significantly higher, the P-gp expressions were significantly decreased after 3 and 6 weeks of treatment, and the Kaplan-Meier survival curves of PFS were significantly longer in the intravenous and intratumor groups（P〈0.05 or P〈0.01）, and the intratumor group showed better effects than the intravenous group（P〈0.05 or P〈0.01）. Compared with the control group, the occurrences of rash, nausea and leukocytopenia were significantly decreased in the intravenous and intratumor groups（P〈0.05）, but without significant difference between the intravenous and intratumor groups（P〉0.05）. Conclusion：Aidi Injection not only improves the efficacy of NP regime, but also has the function of reducing adverse events and preventing against overexpression of P-gp induced by chemotherapy of NP regimen.

Doxorubicin-loaded zein in situ gel for interstitial chemotherapy of colorectal cancer

The aim of this research was to evaluate doxorubicin(DOX)-loaded zein in situ gels,a new drug delivery system in which a liquid state drug can be transformed into semi-solid after intratumoral injection.In vitro release of DOX-loaded zein was investigated and the pharmacokinetics,biodistribution and therapeutic efficacy of these DOX-loaded zein formulations were investigated using BAI B/c nude tumor-bearing mice.In vitro release of DOX from the gels extended up to 7 days.Efficient accumulation of DOX in the tumor with lower drug concentration in blood and normal organs was obtained resulting in effective inhibition of tumor growth and fewer off-target side effects.In conclusion,a DOX-loaded in situ gel was developed with sustained release,enhanced anti-cancer efficacy for colorectal cancer in vivo,and especially with reduced off-target side effects.